Methods for treating, diagnosing, and monitoring rheumatoid arthritis

ABSTRACT

Methods of identifying, diagnosing, and prognosing rheumatoid arthritis are provided, as well as methods of treating rheumatoid arthritis. Also provided are methods for identifying effective rheumatoid arthritis therapeutic agents and predicting responsiveness to rheumatoid arthritis therapeutic agents.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. patent application Ser. No. 12/874,972, filed Sep. 2, 2010, which claims the benefit of priority of provisional U.S. Application No. 61/275,948, filed Sep. 3, 2009 and provisional U.S. Application No. 61/252,424, filed Oct. 16, 2009, the contents of which are hereby incorporated by reference in their entirety.

FIELD

Methods of identifying, diagnosing, and prognosing rheumatoid arthritis are provided, as well as methods of treating rheumatoid arthritis. Also provided are methods for identifying effective rheumatoid arthritis therapeutic agents and predicting responsiveness to rheumatoid arthritis therapeutic agents.

BACKGROUND

Rheumatoid arthritis (RA) is a clinically important, chronic systemic autoimmune inflammatory disease affecting between 1.3 and 2.1 million persons in the United States (See, e.g., Alamanosa and Drosos, Autoimmun. Rev., 4:130-136 (2005)). RA is an autoimmune disorder of unknown etiology. Most RA patients suffer a chronic course of disease that, even with currently available therapies, may result in progressive joint destruction, deformity, disability and even premature death. More than 9 million physician visits and more than 250,000 hospitalizations per year result from RA.

Diagnosis of RA typically relies on clinical and laboratory evaluation of a patient's signs and symptoms. Generally, laboratory evaluation of a patient suspected of having RA may include determination of the level of certain antibodies in serum known as rheumatoid factor (RF) and antibodies to cyclic citrullinated peptide (anti-CCP). (See, e.g., Schellekens et al., Arthritis Rheum., 43:155-163 (2000); DiFranco et al., Rev. Rheum. Engl. Ed., 66(5):251-255 (1999); Rantapaa-Dahlqvist et al., Arthritis Rheum., 48:2741-2749 (2003); Li et al., Bioinformatics 22(12):1503-1507 (2006); Russell et al., J. Rhematol., 33(7):1240-1242 (2006); Ota, Rinsho byori. Jap. J. Clin. Pathol., 54(8)861-868 (2006); Avouac et al., Ann. Rheum. Dis., 65(7):845-851 (2006)). While these antibodies are often found in the serum of RA patients, not all RA patients have them. An additional blood test known as the erythrocyte sedimentation rate (ESR) may also be used. An elevated ESR indicates the general presence of an inflammatory process, although not necessarily RA. Further blood tests may be used to assess the level of other factors, such as C-reactive protein (CRP) that have been associated with RA. In addition, radiographic analysis of affected joints may be performed. In sum, such currently available laboratory tests to diagnose RA are imprecise and imperfect.

In certain instances, diagnosis of RA is made if a patient satisfies certain American College of Rheumatology (ACR) criteria. Certain such criteria include morning stiffness in and around the joints lasting for at least 1 hour before maximal improvement; arthritis of three or more joint areas: at least three joint areas have simultaneously had soft tissue swelling or fluid (not bony overgrowth alone) observed by a physician; the 14 possible joint areas (right and left) are proximal interphalangeal (PIP), metacarpophalangeal (MCP), wrist, elbow, knee, ankle, and metatarsophalangeal (MTP) joints; arthritis of hand joints: at least one joint area swollen as above in wrist, MCP, or PIP joint; symmetric arthritis: simultaneous involvement of the same joint areas (as in arthritis of three or more joint areas, above) on both sides of the body (bilateral involvement of PIP, MCP, or MTP joints is acceptable without absolute symmetry); rheumatoid nodules: subcutaneous nodules over bony prominences or extensor surfaces or in juxta-articular regions that are observed by a physician; serum rheumatoid factor: demonstration of abnormal amounts of serum rheumatoid factor by any method that has been positive in fewer than five percent of normal control patients; radiographic changes: radiographic changes typical of rheumatoid arthritis on posteroanterior hand and wrist X-rays, which must include erosions or unequivocal bony decalcification localized to or most marked adjacent to the involved joints (osteoarthritis changes alone do not qualify). Diagnosis of RA is typically made if a patient satisfies at least four of the above criteria.

A number of published studies report the attempted identification of reliable biomarkers for diagnostic and prognostic purposes. (See e.g., Rioja et al., Arthritis and Rheum. 58(8):2257-2267 (2008); Pyrpasopoulou et al., Mol. Diagn. Ther. 14(1):43-48 (2010); US 2004/0009479; US 2007/0105133; WO 2007/038501; WO 2007/135568; WO 2008/104608; WO 2008/056198; WO 2008/132176; and WO 2008/154423). No clinically validated diagnostic markers, however, e.g., biomarkers, have been identified that enable clinicians or others to accurately define pathophysiological aspects of rheumatoid arthritis, clinical activity, response to therapy, prognosis, or risk of developing the disease. Accordingly, as RA patients seek treatment, there is considerable trial and error involved in the search for therapeutic agent(s) effective for a particular patient. Such trial and error often involves considerable risk and discomfort the patient in order to find the most effective therapy. Thus, there is a need for more effective means for determining which patients will respond to which treatment and for incorporating such determinations into more effective treatment regimens for RA patients.

It would therefore be highly advantageous to have additional diagnostic methods, including molecular-based diagnostic methods, that can be used to objectively identify the presence of and/or classify the disease in a patient, define pathophysiologic aspects of rheumatoid arthritis, clinical activity, response to therapy, including response to treatment with various RA therapeutic agents, prognosis, and/or risk of developing rheumatoid arthritis. In addition, it would be advantageous to have molecular-based diagnostic markers associated with various clinical and/or pathophysiological and/or other biological indicators of disease. Thus, there is a continuing need to identify new molecular biomarkers associated with rheumatoid arthritis as well as other autoimmune disorders. Such associations would greatly benefit the identification of the presence of rheumatoid arthritis in patients or the determination of susceptibility to develop the disease. Such associations would also benefit the identification of pathophysiologic aspects of RA, clinical activity, response to therapy, or prognosis. In addition, statistically and biologically significant and reproducible information regarding such associations could be utilized as an integral component in efforts to identify specific subsets of patients who would be expected to significantly benefit from treatment with a particular therapeutic agent, for example where the therapeutic agent is or has been shown in clinical studies to be of therapeutic benefit in such specific RA patient subpopulation.

The invention described herein meets the above-described needs and provides other benefits.

All references cited herein, including patent applications and publications, are incorporated by reference in their entirety for any purpose.

SUMMARY

The compositions and methods of the invention are based, at least in part, on the definition of four new and distinct molecular phenotypes (also referred to herein as molecular subtypes) of rheumatoid arthritis (RA). These four RA molecular subtypes described herein were defined based on differential gene expression between the subtypes and significant associations of each of the molecular subtypes with certain histology indicators of joint pathology as well as certain biological pathways. The terms “molecular phenotype” and “molecular subtype” are used interchangeably herein.

Accordingly, in one aspect, therapeutic targets for the treatment of a certain molecular subtype of RA, described herein as lymphoid-rich (L) subtype, are provided. In certain embodiments, a L subtype therapeutic target is selected from one or a combination of genes listed in Table 5. In certain embodiments, a L subtype therapeutic target is selected from one or a combination of genes listed in Table 1. In certain embodiments, a L subtype therapeutic target is selected from one or a combination of genes listed in Table 10. In certain embodiments, a L subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 5. In certain embodiments, a L subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 1. In certain embodiments, a L subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 10. In certain embodiments, a therapeutic target of L subtype of RA is selected from one or more of CD20 (synonymous with MS4A1), CTLA4, CD3, CRTAM, IL2Rβ, IL2Rγ, CD19, HLAII, CD79a, CD79b, FcRH5 (synonymous with IRTA2), CD38, IL21R, IL12Rβ1, and IL12Rβ2.

In another aspect, methods of diagnosing a certain subtype of RA, described herein as L subtype, comprise measuring the gene expression of one or a combination of genes listed in Table 5, or measuring the amount of protein expressed by one or a combination of genes listed in Table 5. In certain embodiments, one or more of the genes identified in Table 5, or proteins encoded by said genes, are biomarkers of the L subtype. In certain embodiments, methods of diagnosing L subtype RA comprise measuring the gene expression of one or a combination of genes listed in Table 1, or measuring the amount of protein expressed by one or a combination of genes listed in Table 1. In certain embodiments, one or more of the genes identified in Table 1, or proteins encoded by said genes, are biomarkers of the L subtype. In certain embodiments, methods of diagnosing L subtype RA comprise measuring the gene expression of one or a combination of genes listed in Table 10, or measuring the amount of protein expressed by one or a combination of genes listed in Table 10. In certain embodiments, one or more of the genes identified in Table 10, or proteins encoded by said genes, are biomarkers of the L subtype. In certain embodiments, methods of diagnosing L subtype of RA comprise measuring the gene expression or protein expression of one or more of CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, methods of diagnosing L subtype of RA comprise measuring protein expression of CXCL13 and/or sFcRH5 and/or RF in serum. In certain embodiments, a patient is diagnosed with L subtype RA when the serum level of CXCL13 is greater than 116.6 pg/ml, or greater than 150 pg/ml, or greater than 200 pg/ml, or greater than 250 pg/ml, or greater than 300 pg/ml. In certain embodiments, a patient is diagnosed with L subtype RA when the serum level of sFcRH5 is greater than 126.7 ng/ml, or greater than 150 ng/ml, or greater than 200 ng/ml, or greater than 250 ng/ml, or greater than 300 ng/ml. In certain embodiments, a patient is diagnosed with L subtype RA when the serum is positive for RF and when the serum level of sFcRH5 is elevated compared to a control sample. In certain such embodiments, the serum level of sFcRH5 is greater than 126.7 ng/ml, or greater than 150 ng/ml, or greater than 200 ng/ml, or greater than 250 ng/ml, or greater than 300 ng/ml. In certain embodiments, a patient is diagnosed with L subtype RA when the serum is positive for RF and when the serum level of both sFcRH5 and CXCL13 are elevated compared to a control sample. In certain such embodiments, the serum level of sFcRH5 is greater than 126.7 ng/ml, or greater than 150 ng/ml, or greater than 200 ng/ml, or greater than 250 ng/ml, or greater than 300 ng/ml and the serum level of CXCL13 is greater than 116.6 pg/ml, or greater than 150 pg/ml, or greater than 200 pg/ml, or greater than 250 pg/ml, or greater than 300 pg/ml.

In another aspect, therapeutic targets for the treatment of a certain molecular subtype of RA, described herein as myeloid-rich (M) subtype, are provided. In certain embodiments, a M subtype therapeutic target is selected from one or a combination of genes listed in Table 6. In certain embodiments, a M subtype therapeutic target is selected from one or a combination of genes listed in Table 2. In certain embodiments, a M subtype therapeutic target is selected from one or a combination of genes listed in Table 11. In certain embodiments, a M subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 6. In certain embodiments, a M subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 2. In certain embodiments, a M subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 11. In certain embodiments, a therapeutic target of M subtype of RA is selected from one or more of CLEC5A, CLEC7A, ALCAM, IL1RAP, IRAK1, NRP2, TREM1, and VEGF.

In another aspect, methods of diagnosing a certain subtype of RA, described herein as the M subtype, comprise measuring the gene expression of one or a combination of genes listed in Table 6, or measuring the amount of protein expressed by one or a combination of genes listed in Table 6. In certain embodiments, one or more of the genes identified in Table 6, or proteins encoded by said genes, are biomarkers of the M subtype. In certain embodiments, methods of diagnosing M subtype RA comprise measuring the gene expression of one or a combination of genes listed in Table 2, or measuring the protein expressed by one or a combination of genes listed in Table 2. In certain embodiments, one or more of the genes identified in Table 2, or proteins encoded by said genes, are biomarkers of the M subtype. In certain embodiments, methods of diagnosing M subtype RA comprise measuring the gene expression of one or a combination of genes listed in Table 11, or measuring the protein expressed by one or a combination of genes listed in Table 11. In certain embodiments, one or more of the genes identified in Table 11, or proteins encoded by said genes, are biomarkers of the M subtype. In certain embodiments, methods of diagnosing M subtype of RA comprise measuring the gene expression or protein expression of one or more of ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11.

In another aspect, therapeutic targets for the treatment of a certain molecular subtype of RA, described herein as fibroblast-rich type 2 (F2) subtype, are provided. In certain embodiments, a F2 subtype therapeutic target is selected from one or a combination of genes listed in Table 7. In certain embodiments, a F2 subtype therapeutic target is selected from one or a combination of genes listed in Table 3. In certain embodiments, a F2 subtype therapeutic target is selected from one or a combination of genes listed in Table 12. In certain embodiments, a F2 subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 7. In certain embodiments, a F2 subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 3. In certain embodiments, a F2 subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 12. In certain embodiments, a therapeutic target of F2 subtype of RA is selected from one or more of IL17D, IL17RC, TIMP3, and TNFRSF11B.

In another aspect, methods of diagnosing a certain subtype of RA, described herein as the F2 subtype, comprise measuring the gene expression of one or a combination of genes listed in Table 7, or measuring the protein expressed by one or a combination of genes listed in Table 7. In certain embodiments, one or more of the genes identified in Table 7, or proteins encoded by said genes, are biomarkers of the F2 subtype. In certain embodiments, methods of diagnosing F2 subtype RA comprise measuring the gene expression of one or a combination of genes listed in Table 3, or measuring the protein expressed by one or a combination of genes listed in Table 3. In certain embodiments, one or more of the genes identified in Table 3, or proteins encoded by said genes, are biomarkers of the F2 subtype. In certain embodiments, methods of diagnosing F2 subtype RA comprise measuring the gene expression of one or a combination of genes listed in Table 12, or measuring the protein expressed by one or a combination of genes listed in Table 12. In certain embodiments, one or more of the genes identified in Table 12, or proteins encoded by said genes, are biomarkers of the F2 subtype. In certain embodiments, methods of diagnosing F2 subtype of RA comprise measuring the gene expression or protein expression of one or more of FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D.

In another aspect, therapeutic targets for the treatment of a certain molecular subtype of RA, described herein as fibroblast-rich type 1 (F1) subtype, are provided. In certain embodiments, a F1 subtype therapeutic target is selected from one or a combination of genes listed in Table 8. In certain embodiments, a F1 subtype therapeutic target is selected from one or a combination of genes listed in Table 4. In certain embodiments, a F1 subtype therapeutic target is selected from one or a combination of genes listed in Table 13. In certain embodiments, a F1 subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 8. In certain embodiments, a F1 subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 4. In certain embodiments, a F1 subtype therapeutic target is selected from one or a combination of proteins encoded by one or a combination of genes listed in Table 13. In certain embodiments, a therapeutic target of F1 subtype of RA is selected from one or more of CDH11, ITGA11, and CLEC11A.

In another aspect, methods of diagnosing a certain subtype of RA, described herein as the F1 subtype, comprise measuring the gene expression of one or a combination of genes listed in Table 8, or measuring the protein expressed by one or a combination of genes listed in Table 8. In certain embodiments, one or more of the genes identified in Table 8, or proteins encoded by said genes, are biomarkers of the F1 subtype. In certain embodiments, methods of diagnosing F1 subtype RA comprises measuring the gene expression of one or a combination of genes listed in Table 4, or measuring the protein expressed by one or a combination of genes listed in Table 4. In certain embodiments, one or more of the genes identified in Table 4, or proteins encoded by said genes, are biomarkers of the F1 subtype. In certain embodiments, methods of diagnosing F1 subtype RA comprises measuring the gene expression of one or a combination of genes listed in Table 13, or measuring the protein expressed by one or, a combination of genes listed in Table 13. In certain embodiments, one or more of the genes identified in Table 13, or proteins encoded by said genes, are biomarkers of the F1 subtype. In certain embodiments, methods of diagnosing F1 subtype of RA comprise measuring the gene expression or protein expression of one or more of ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF.

In one aspect, gene expression is measured by microarray. In another aspect gene expression is measured by real-time quantitative polymerase chain reaction (qPCR). In another aspect, gene expression is measured by multiplex-PCR. According to another embodiment, gene expression is measured by observing protein expression levels of an aforementioned gene. According to another embodiment, expression of a gene of interest is considered elevated when compared to a healthy control if the relative mRNA level of the gene of interest is greater than 2 fold of the level of a control gene mRNA. According to another embodiment, the relative mRNA level of the gene of interest is greater than 3 fold, fold, 10 fold, 15 fold, 20 fold, 25 fold, or 30 fold compared to a healthy control gene expression level. In one aspect, the gene expression level is measured by a method selected from a PCR method, a microarray method, or an immunoassay method. In one embodiment, the microarray method comprises the use of a microarray chip having one or more nucleic acid molecules that can hybridize under stringent conditions to a nucleic acid molecule encoding a gene mentioned above or having one or more polypeptides (such as peptides or antibodies) that can bind to one or more of the proteins encoded by the genes mentioned above. In one embodiment, the PCR method is qPCR. In one embodiment, the PCR method is multiplex-PCR. According to one embodiment, the immunoassay method comprises binding an antibody to protein expressed from a gene mentioned above in a patient sample and determining if the protein level from the patient sample is elevated. In certain embodiments, the immunoassay method is an enzyme-linked immunosorbent assay (ELISA). In certain embodiments, protein expression of CXCL13, sFcRH5, and/or RF are measured by ELISA.

In one aspect, a method of identifying a subtype of rheumatoid arthritis in a subject is provided, the method comprising measuring in a biological sample obtained from the subject the expression of one or more genes, or one or more proteins encoded by said genes, associated with a certain subtype. In one aspect, the subtype of RA is selected from L subtype, M subtype, F2 subtype, and F1 subtype as described herein. In certain embodiments, the subtype of RA is L subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10. In certain embodiments, the subtype of RA is L subtype and the one or more genes are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10 and the expression of the one or more genes is measured using the corresponding probes listed in Table 1 or Table 5 or Table 10, respectively. In certain embodiments, the subtype of RA is L subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, the biological sample is a serum sample, and the protein expression measured is selected from CXCL13 and sFcRH5. In certain embodiments, the biological sample is a serum sample, the serum sample is positive for RF, and the protein expression measured is selected from CXCL13 and sFcRH5. In certain embodiments, the biological sample is a serum sample, the serum sample is positive for RF, and the protein expression measured is CXCL13 and sFcRH5. In certain embodiments, the subtype of RA is identified as L subtype when the serum level of CXCL13 is greater than 116.6 pg/ml, or greater than 150 pg/ml, or greater than 200 pg/ml, or greater than 250 pg/ml, or greater than 300 pg/ml. In certain embodiments, the subtype of RA is identified as L subtype when the serum level of FcRH5 is greater than 126.7 ng/ml, or greater than 150 ng/ml, or greater than 200 ng/ml, or greater than 250 ng/ml, or greater than 300 ng/ml. In certain embodiments, the subtype of RA is M subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11. In certain embodiments, the subtype of RA is M subtype and the one or more genes are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11 and the expression of the one or more genes is measured using the corresponding probes listed in Table 2 or Table 6 or Table 11, respectively. In certain embodiments, the subtype of RA is M subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11. In certain embodiments, the subtype of RA is F2 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in. Table 3 or Table 7 or Table 12. In certain embodiments, the subtype of RA is F2 subtype and the one or more genes are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12 and the expression of the one or more genes is measured using the corresponding probes listed in Table 3 or Table 7 or Table 12, respectively. In certain embodiments, the subtype of RA is F2 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D. In certain embodiments, the subtype of RA is F1 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13. In certain embodiments, the subtype of RA is F1 subtype and the one or more genes are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13 and the expression of the one or more genes is measured using the corresponding probes listed in Table 4 or Table 8 or Table 13, respectively. In certain embodiments, the subtype of RA is F1 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF.

In another aspect, a method for predicting whether a subject with RA will respond to a RA therapeutic agent is provided, the method comprising measuring in a biological sample obtained from the subject the expression of one or more genes of a gene signature, or the expression of one or more proteins encoded by said genes (a protein signature), associated with a molecular subtype of RA. In one aspect, the gene signature or protein signature is associated with a molecular subtype of RA selected from L subtype, M subtype, F2 subtype, and F1 subtype as described herein. In certain embodiments, the gene signature or protein signature is associated with L subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10. In certain embodiments, the gene signature is associated with L subtype and the one or more genes are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10 and the expression of the one or more genes is measured using the corresponding probes listed in Table 1 or Table 5 or Table 10, respectively. In certain embodiments, gene signature or protein signature is associated with L subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, the protein signature is associated with L subtype and one or more proteins encoded by said genes are selected from CXCL13, sFcRH5 and RF. In certain embodiments, the biological sample is a serum sample. In certain embodiments, the RA therapeutic agent is a B-cell antagonist. In certain embodiments, the B-cell antagonist is selected from CD22 antibodies, CD20 antibodies, BR3 antibodies, and BR3-Fc immunoadhesins. In certain embodiments, the CD20 antibody is selected from rituximab, ibritumomab tiuxetan, tositumomab, 1F5, 2H7, and A20. In certain embodiments, methods for predicting whether a subject with RA will respond to rituximab are provided, comprising measuring serum levels of CXCL13, sFcRH5, and/or RF. In one embodiment, a subject with RA is predicted to respond to rituximab when the serum level of CXCL13 is greater than 116.6 pg/ml. In one embodiment, a subject with RA is predicted to respond to rituximab when the serum level of sFcRH5 is greater than 126.7 ng/ml. In one embodiment, a subject with RA is predicted to respond to rituximab when the serum level of CXCL13 is greater than 116.6 pg/ml and the serum level of sFcRH5 is greater than 126.7 ng/ml. In one embodiment, a subject with RA is predicted to respond to rituximab when the serum is positive for RF and the serum level of CXCL13 is greater than 116.6 pg/ml and the serum level of sFcRH5 is greater than 126.7 ng/ml.

In another aspect, the gene signature or protein signature mentioned above is associated with M subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11. In certain embodiments, the gene signature is associated with M subtype and the one or more genes are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11 and the expression of the one or more genes is measured using the corresponding probes listed in Table 2 or Table 6 or Table 11, respectively. In certain embodiments, the gene signature or protein signature is associated with M subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11.

In yet another aspect, the gene signature or protein signature mentioned above is associated with F2 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12. In certain embodiments, the gene signature is associated with F2 subtype and the one or more genes are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12 and the expression of the one or more genes is measured using the corresponding probes listed in Table 3 or Table 7 or Table 12, respectively. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D.

In still another aspect, the gene signature or protein signature is associated with F1 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13. In certain embodiments, the gene signature is associated with F1 subtype and the one or more genes are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13 and the expression of the one or more genes is measured using the corresponding probes listed in Table 4 or Table 8 or Table 13, respectively. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, T1141, and VWF.

In certain embodiments, the RA therapeutic agent targets a biological pathway selected from cytokine/chemokine, lymphocyte, dendritic cell, macrophage, fibroblast, osteoblast and osteoclast. In certain embodiments, the RA therapeutic agent is selected from a TNFα inhibitor, a B-cell antagonist, an IL-17A/F binding agent, an IL-6 binding agent, an inhibitor of costimulation, e.g., an inhibitor of the CD28/B7 pathway, a CD4 binding agent. In certain embodiments, the inhibitor of the CD28/B7 pathway is CTLA4-Ig.

In yet another aspect, a method of diagnosing or prognosing RA in a subject is provided, the method comprising measuring in a biological sample obtained from the subject the expression of one or more genes, or one or more proteins encoded by said genes, associated with a certain subtype. In one aspect, the subtype of RA is selected from L subtype, M subtype, F2 subtype, and F1 subtype as described herein. In certain embodiments, the subtype of RA is L subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10. In certain embodiments, the subtype of RA is. L subtype and the one or more genes are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10 and the expression of the one or more genes is measured using the corresponding probes listed in Table 1 or Table 5 or Table 10, respectively. In certain embodiments, the subtype of RA is L subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, the methods comprise measuring in a serum sample obtained from the subject, the protein expression of CXCL13, sFcRH5, and/or RF. In certain embodiments, a patient is diagnosed or prognosed with L subtype RA when the serum level of CXCL13 is greater than 116.6 pg/ml, or greater than 150 pg/ml, or greater than 200 pg/ml, or greater than 250 pg/ml, or greater than 300 pg/ml. In certain embodiments, a patient is diagnosed or prognosed with L subtype RA when the serum level of FcRH5 is greater than 126.7 ng/ml, or greater than 150 ng/ml, or greater than 200 ng/ml, or greater than 250 ng/ml, or greater than 300 ng/ml. In certain embodiments, the biological sample is a serum sample, the serum sample is positive for RF, and the protein expression measured is selected from one of CXCL13 and sFcRH5. In certain embodiments, the biological sample is a serum sample, the serum sample is positive for RF, and the protein expression measured is both of CXCL13 and sFcRH5. In certain embodiments, the subtype of RA is M subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11. In certain embodiments, the subtype of RA is M subtype and the one or more genes are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11 and the expression of the one or more genes is measured using the corresponding probes listed in Table 2 or Table 6 or Table 11, respectively. In certain embodiments, the subtype of RA is M subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11. In certain embodiments, the subtype of RA is F2 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12. In certain embodiments, the subtype of RA is F2 subtype and the one or more genes are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12 and the expression of the one or more genes is measured using the corresponding probes listed in Table 3 or Table 7 or Table 12, respectively. In certain embodiments, the subtype of RA is F2 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D. In certain embodiments, the subtype of RA is F1 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13. In certain embodiments, the subtype of RA is F1 subtype and the one or more genes are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13 and the expression of the one or more genes is measured using the corresponding probes listed in Table 4 or Table 8 or Table 13, respectively. In certain embodiments, the subtype of RA is F1 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF.

In a still further aspect, a method of aiding in the diagnosis or prognosis of RA in a subject is provided, the method comprising measuring in a biological sample obtained from the subject the expression of one or more genes, or one or more proteins encoded by said genes, associated with a given subtype. In one aspect, the subtype of RA is selected from L subtype, M subtype, F2 subtype, and F1 subtype as described herein. In certain embodiments, the subtype of RA is L subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10. In certain embodiments, the subtype of RA is L subtype and the one or more genes are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10 and the expression of the one or more genes is measured using the corresponding probes listed in Table 1 or Table 5 or Table 10, respectively. In certain embodiments, the subtype of RA is L subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, the methods comprise measuring in a serum sample obtained from the subject, the protein expression of CXCL13, sFcRH5, and/or RF. In certain embodiments, diagnosis or prognosis of L subtype RA is aided when the serum level of CXCL13 is greater than 116.6 pg/ml, or greater than 150 pg/ml, or greater than 200 pg/ml, or greater than 250 pg/ml, or greater than 300 pg/ml. In certain embodiments, diagnosis or prognosis of L subtype RA is aided when the serum level of FcRH5 is greater than 126.7 ng/ml, or greater than 150 ng/ml, or greater than 200 ng/ml, or greater than 250 ng/ml, or greater than 300 ng/ml. In certain embodiments, the biological sample is a serum sample, the serum sample is positive for RF, and the protein expression measured is selected from one of CXCL13 and sFcRH5. In certain embodiments, the biological sample is a serum sample, the serum sample is positive for RF, and the protein expression measured is both of CXCL13 and sFcRH5. In certain embodiments, the subtype of RA is M subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11. In certain embodiments, the subtype of RA is M subtype and the one or more genes are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11 and the expression of the one or more genes is measured using the corresponding probes listed in Table 2 or Table 6 or Table 11, respectively. In certain embodiments, the subtype of RA is M subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11. In certain embodiments, the subtype of RA is F2 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12. In certain embodiments, the subtype of RA is F2 subtype and the one or more genes are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12 and the expression of the one or more genes is measured using the corresponding probes listed in Table 3 or Table 7 or Table 12, respectively. In certain embodiments, the subtype of RA is F2 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D. In certain embodiments, the subtype of RA is F1 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13. In certain embodiments, the subtype of RA is F1 subtype and the one or more genes are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13 and the expression of the one or more genes is measured using the corresponding probes listed in Table 4 or Table 8 or Table 13, respectively. In certain embodiments, the subtype of RA is F1 subtype and the one or more genes, or one or more proteins encoded by said genes, are selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF.

In one aspect, a method of treating RA in a subject in whom a gene signature or a protein signature associated with a molecular subtype of RA has been detected. In one aspect, the gene signature or protein signature is associated with a molecular subtype of RA selected from L subtype, M subtype, F2 subtype, and F1 subtype as described herein. In certain embodiments, the gene signature is associated with L subtype and the gene signature comprises one or a combination of genes listed in Table 1 or Table 5 or Table 10. In certain embodiments, the gene signature is associated with L subtype and the one or more genes are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10 and the expression of the one or more genes is measured using the corresponding probes listed in Table 1 or Table 5 or Table 10, respectively. In certain embodiments, the gene signature is associated with L subtype and the gene signature comprises one or a combination of genes selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, a protein signature is associated with L subtype and the protein signature comprises one or a combination of proteins selected from CXCL13, sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, the protein signature comprises CXCL13, sFcRH5, and/or RF. In certain embodiments, the gene signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 2 or Table 6 or Table 11. In certain embodiments, the gene signature is associated with M subtype and the one or more genes are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11 and the expression of the one or more genes is measured using the corresponding probes listed in Table 2 or Table 6 or Table 11, respectively. In certain embodiments, the gene signature or protein signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 3 or Table 7 or Table 12. In certain embodiments, the gene signature is associated with F2 subtype and the one or more genes are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12 and the expression of the one or more genes is measured using the corresponding probes listed in Table 3 or Table 7 or Table 12, respectively. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 4 or Table 8 or Table 13. In certain embodiments, the gene signature is associated with F1 subtype and the one or more genes are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13 and the expression of the one or more genes is measured using the corresponding probes listed in Table 4 or Table 8 or Table 13, respectively. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF.

In another aspect, a method of treating a subject having a molecular subtype of RA is provided, the method comprising administering to the subject a therapeutic agent effective to treat the subtype in a subject in whom a gene signature or a protein signature associated with the molecular subtype of RA has been detected. In one aspect, the gene signature or protein signature is associated with a molecular subtype of RA selected from L subtype, M subtype, F2 subtype, and F1 subtype as described herein. In certain embodiments, the gene signature is associated with L subtype and the gene signature comprises one or a combination of genes listed in Table 1 or Table 5 or Table 10. In certain embodiments, the gene signature is associated with L subtype and the one or more genes are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10 and the expression of the one or more genes is measured using the corresponding probes listed in Table 1 or Table 5 or Table 10, respectively. In certain embodiments, the gene signature is associated with L subtype and the gene signature comprises one or a combination of genes selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, a protein signature is associated with L subtype and the protein signature comprises one or a combination of proteins selected from CXCL13, sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, the protein signature comprises CXCL13, sFcRH5 and/or RF. In certain embodiments, the gene signature or protein signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 2 or Table 6 or Table 11. In certain embodiments, the gene signature is associated with M subtype and the one or more genes are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11 and the expression of the one or more genes is measured using the corresponding probes listed in Table 2 or Table 6 or Table 11, respectively. In certain embodiments, the gene signature or protein signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, MMP12, CCL2, VEGFA, and S100A11. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 3 or Table 7 or Table 12. In certain embodiments, the gene signature is associated with F2 subtype and the one or more genes are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12 and the expression of the one or more genes is measured using the corresponding probes listed in Table 3 or Table 7 or Table 12, respectively. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, FZD10, FZD7, FZD8, and IL17D. In certain embodiments, the gene signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes listed in Table 4 or Table 8 or Table 13. In certain embodiments, the gene signature is associated with F1 subtype and the one or more genes are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13 and the expression of the one or more genes is measured using the corresponding probes listed in Table 4 or Table 8 or Table 13, respectively. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF.

In another aspect, a method comprising manufacturing a RA therapeutic agent is provided, which includes packaging the agent with instructions to administer the agent to a subject who has or is believed to have RA and in whom a gene signature or a protein signature associated with a molecular subtype of RA has been detected. In one aspect, the gene signature or protein signature is associated with a molecular subtype of RA selected from L subtype, M subtype, F2 subtype, and F1 subtype as described herein. In certain embodiments, the gene signature is associated with L subtype and the gene signature comprises one or a combination of genes listed in Table 1 or Table 5 or Table 10. In certain embodiments, the gene signature is associated with L subtype and the one or more genes are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10 and the expression of the one or more genes is measured using the corresponding probes listed in Table 1 or Table 5 or Table 10, respectively. In certain embodiments, gene signature is associated with L subtype and the gene signature comprises one or a combination of genes selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, a protein signature is associated with L subtype and the protein signature comprises one or a combination of proteins selected from CXCL13, sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, the protein signature comprises CXCL13, sFcRH5 and/or RF. In certain embodiments, the gene signature or protein signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 2 or Table 6 or Table 11. In certain embodiments, the gene signature is associated with M subtype and the one or more genes are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11 and the expression of the one or more genes is measured using the corresponding probes listed in Table 2 or Table 6 or Table 11, respectively. In certain embodiments, the gene signature or protein signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 3 or Table 7 or Table 12. In certain embodiments, the gene signature is associated with F2 subtype and the one or more genes are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12 and the expression of the one or more genes is measured using the corresponding probes listed in Table 3 or Table 7 or Table 12, respectively. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 4 or Table 8 or Table 13. In certain embodiments, the gene signature is associated with F1 subtype and the one or more genes are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13 and the expression of the one or more genes is measured using the corresponding probes listed in Table 4 or Table 8 or Table 13, respectively. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF.

In one aspect, a method for selecting a patient suffering from RA for treatment with a RA therapeutic agent is provided, the method comprising detecting the presence of a gene signature or protein signature associated with a molecular subtype of RA. In certain embodiments, the gene signature is associated with L subtype and the gene signature comprises one or a combination of genes listed in Table 1 or Table 5 or Table 10. In certain embodiments, the gene signature is associated with L subtype and the one or more genes are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10 and the expression of the one or more genes is measured using the corresponding probes listed in Table 1 or Table 5 or Table 10, respectively. In certain embodiments, gene signature is associated with L subtype and the gene signature comprises one or a combination of genes selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, a protein signature is associated with L subtype and the protein signature comprises one or a combination of proteins selected from CXCL13, sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, the protein signature comprises CXCL13, sFcRH5, and/or RF. In certain embodiments, the gene signature or protein signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 2 or Table 6 or Table 11. In certain embodiments, the gene signature is associated with M subtype and the one or more genes are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11 and the expression of the one or more genes is measured using the corresponding probes listed in Table 2 or Table 6 or Table 11, respectively. In certain embodiments, the gene signature or protein signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 3 or Table 7 or Table 12. In certain embodiments, the gene signature is associated with F2 subtype and the one or more genes are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12 and the expression of the one or more genes is measured using the corresponding probes listed in Table 3 or Table 7 or Table 12, respectively. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 4 or Table 8 or Table 13. In certain embodiments, the gene signature is associated with F1 subtype and the one or more genes are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13 and the expression of the one or more genes is measured using the corresponding probes listed in Table 4 or Table 8 or Table 13, respectively. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF.

In another aspect, a method of assessing a stage of RA in a subject or in a sample obtained from the subject is provided, the method comprising detecting in a biological sample obtained from the subject, the presence of a gene signature or protein signature associated with a molecular subtype of RA. In certain embodiments, the gene signature or protein signature is associated with L subtype and the gene signature comprises one or a combination of genes listed in Table 1 or Table 5. In certain embodiments, the gene signature is associated with L subtype and the one or more genes are selected from one or a combination of genes listed in Table 1 or Table 5 or Table 10 and the expression of the one or more genes is measured using the corresponding probes listed in Table 1 or Table 5 or Table 10, respectively. In certain embodiments, gene signature is associated with L subtype and the gene signature comprises one or a combination of genes selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, a protein signature is associated with L subtype and the protein signature comprises one or a combination of proteins selected from CXCL13, sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1. In certain embodiments, the protein signature comprises CXCL13, sFcRH5, and/or RF. In certain embodiments, the gene signature or protein signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 2 or Table 6 or Table 11. In certain embodiments, the gene signature is associated with M subtype and the one or more genes are selected from one or a combination of genes listed in Table 2 or Table 6 or Table 11 and the expression of the one or more genes is measured using the corresponding probes listed in Table 2 or Table 6 or Table 11, respectively. In certain embodiments, the gene signature or protein signature is associated with M subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 3 or Table 7 or Table 12. In certain embodiments, the gene signature is associated with F2 subtype and the one or more genes are selected from one or a combination of genes listed in Table 3 or Table 7 or Table 12 and the expression of the one or more genes is measured using the corresponding probes listed in Table 3 or Table 7 or Table 12, respectively. In certain embodiments, the gene signature or protein signature is associated with F2 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, listed in Table 4 or Table 8 or Table 13. In certain embodiments, the gene signature is associated with F1 subtype and the one or more genes are selected from one or a combination of genes listed in Table 4 or Table 8 or Table 13 and the expression of the one or more genes is measured using the corresponding probes listed in Table 4 or Table 8 or Table 13, respectively. In certain embodiments, the gene signature or protein signature is associated with F1 subtype and the gene signature or protein signature comprises one or a combination of genes, or proteins encoded by said genes, selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF.

In yet another aspect, kits for diagnosing a molecular subtype of RA in a patient comprising detecting a gene signature associated with the molecular subtype in a biological sample are provided. In certain embodiments, a kit for diagnosing L subtype is provided and comprises (1) one or more nucleic acid molecules that hybridize with a gene selected from CXCL13, FcRH5 (synonymous with IRTA2), sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1; and (2) instructions for measuring the expression levels of the gene from a RA patient sample, wherein elevated expression levels of any one, combination or all of said genes is indicative of L subtype. In certain embodiments, a kit for diagnosing M subtype is provided and comprises (1) one or more nucleic acid molecules that hybridize with a gene selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11; and (2) instructions for measuring the expression levels of the gene from a RA patient sample, wherein elevated expression levels of any one, combination or all of said genes is indicative of M subtype. In certain embodiments, a kit for diagnosing F2 subtype is provided and comprises (1) one or more nucleic acid molecules that hybridize with a gene selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D; and (2) instructions for measuring the expression levels of the gene from a RA patient sample, wherein elevated expression levels of any one, combination or all of said genes is indicative of F2 subtype. In certain embodiments, a kit for diagnosing F1 subtype is provided and comprises (1) one or more nucleic acid molecules that hybridize with a gene selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF; and (2) instructions for measuring the expression levels of the gene from a RA patient sample, wherein elevated expression levels of any one, combination or all of said genes is indicative of F1 subtype. In certain embodiments, the gene expression level is measured by assaying for mRNA levels. In certain embodiments, the assay comprises a PCR method and/or the use of a microarray chip. In one embodiment, the PCR method is qPCR. In one embodiment, the PCR method is multiplex-PCR. In certain embodiments, kits include at least one enzyme selected from a nuclease, a ligase, and a polymerase.

In a further aspect, kits for diagnosing a molecular subtype of RA in a patient comprising detecting expression of one or more proteins associated with the molecular subtype in a biological sample from the patient are provided. In certain embodiments, a kit for diagnosing L subtype is provided and comprises (1) one or more protein molecules, for example including, but not limited to, antibodies, that bind to a protein selected from CXCL13, sFcRH5 (synonymous with sIRTA2), LTβ, ICAM3, IL18, PACAP, TNFRSF7, IgJ, IGM, IgG, and XBP1; and (2) instructions for measuring the expression levels of the protein from a RA patient sample, wherein elevated expression levels of any one, combination or all of said proteins is indicative of L subtype. In certain embodiments, the proteins detected are selected from CXCL13, sFcRH5, RF and combinations thereof. In certain embodiments, a kit for diagnosing M subtype is provided and comprises (1) one or more protein molecules that bind to a protein selected from ADAM8, CTSB, CXCL3, ICAM1, IL18BP, IL1B, IL8, MMP12, CCL2, VEGFA, and S100A11; and (2) instructions for measuring the expression levels of the protein from a RA patient sample, wherein elevated expression levels of any one, combination or all of said proteins is indicative of M subtype. In certain embodiments, a kit for diagnosing F2 subtype is provided and comprises (1) one or more protein molecules that bind to a protein selected from FGF10, FGF18, FGF2, LRP6, TGFβ2, WNT11, BMP6, BTC, CLU, CRLF1, TIMP3, FZD10, FZD7, FZD8, and IL17D; and (2) instructions for measuring the expression levels of the protein from a RA patient sample, wherein elevated expression levels of any one, combination or all of said proteins is indicative of F2 subtype. In certain embodiments, a kit for diagnosing F1 subtype is provided and comprises (1) one or more protein molecules that bind to a protein selected from ITGA11, MMP11, MMP13, MMP16, MMP28, ADAM12, ADAM22, CTSK, CTHRC1, ENPEP, POSTN, ANGPT2, SFRP2, TIE1, and VWF; and (2) instructions for measuring the expression levels of the protein from a RA patient sample, wherein elevated expression levels of any one, combination or all of said proteins is indicative of F1 subtype. In certain embodiments, the protein molecule is an antibody, a peptide, or a peptibody. In a further embodiment, the kit comprises a microarray chip for detecting the protein molecule(s).

In one aspect, a method of treating rheumatoid arthritis in a patient comprising administering an effective amount of a RA therapeutic agent to the patient to treat the rheumatoid arthritis, provided that a serum sample from the patient contains an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination thereof is provided. In a further embodiment, the serum sample is positive for RF. In certain embodiments, the RA therapeutic agent is a B-cell antagonist. In certain embodiments, the B-cell antagonist is selected from an antibody to CD22, an antibody to CD20, an antibody to BR3, and a BR3-Fc immunoadhesin. In certain embodiments, the B-cell antagonist is an antibody to CD20 and the antibody to CD20 is selected from rituximab, ibritumomab tiuxetan, tositumomab, 1F5, 2H7, and A20.

In another aspect, a method of treating rheumatoid arthritis in a patient comprising administering to the patient an effective amount of a B-cell antagonist, wherein before the administration a serum sample from the patient was determined to contain an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination thereof, whereby the amount or amounts of CXCL13, sFcRH5, or a combination thereof indicates that the patient will respond to treatment with the antagonist is provided. In a further embodiment, the serum sample is positive for RF. In certain embodiments, the RA therapeutic agent is a B-cell antagonist. In certain embodiments, the B-cell antagonist is selected from an antibody to CD22, an antibody to CD20, an antibody to BR3, and a BR3-Fc immunoadhesin. In certain embodiments, the B-cell antagonist is an antibody to CD20 and the antibody to CD20 is selected from rituximab, ibritumomab tiuxetan, tositumomab, 1F5, 2H7, and A20.

In yet another aspect, a method of treating rheumatoid arthritis in a patient comprising administering to the patient an effective amount of a B-cell antagonist, wherein before the administration a serum sample from the patient was determined to contain an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination thereof, whereby the amount or amounts of CXCL13, sFcRH5, or a combination thereof indicates that the patient is likely to respond favorably to treatment with the antagonist is provided. In a further embodiment, the serum sample is positive for RF. In certain embodiments, the RA therapeutic agent is a B-cell antagonist. In certain embodiments, the B-cell antagonist is selected from an antibody to CD22, an antibody to CD20, an antibody to BR3, and a BR3-Fc immunoadhesin. In certain embodiments, the B-cell antagonist is an antibody to CD20 and the antibody to CD20 is selected from rituximab, ibritumomab tiuxetan, tositumomab, 1F5, 2H7, and A20.

In still another aspect, a method for advertising a B-cell antagonist or a pharmaceutically acceptable composition thereof comprising promoting, to a target audience, the use of that antagonist or pharmaceutical composition thereof for treating a patient or patient population with rheumatoid arthritis from which a serum sample has been obtained showing an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination of these amounts is provided. In a further embodiment, the serum sample is positive for RF.

In one aspect, an article of manufacture comprising, packaged together, a pharmaceutical composition comprising a B-cell antagonist and a pharmaceutically acceptable carrier and a label stating that the antagonist or pharmaceutical composition is indicated for treating patients with rheumatoid arthritis which a serum sample has been obtained showing an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination of these amounts is provided. In a further embodiment, the serum sample is positive for RF.

In another aspect, a method for manufacturing a B-cell antagonist or a pharmaceutical composition thereof comprising combining in a package the antagonist or pharmaceutical composition and a label stating that the antagonist or pharmaceutical composition is indicated for treating patients with rheumatoid arthritis from which a serum sample has been obtained showing an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination of these amounts is provided. In a further embodiment, the serum sample is positive for RF.

In yet another aspect, a method of providing a treatment option for patients with rheumatoid arthritis comprising packaging a B-cell antagonist in a vial with a package insert containing instructions to treat patients with rheumatoid arthritis from whom a sample has been obtained that contains an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination of these amounts is provided. In a further embodiment, the sample is positive for RF.

In still another aspect, a method of specifying a B-cell antagonist for use in a rheumatoid arthritis patient subpopulation, the method comprising providing instruction to administer the B-cell antagonist to a patient subpopulation characterized by the presence in a serum sample from said subpopulation of an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination of these amounts is provided. In a further embodiment, the serum sample is positive for RF.

In one aspect, a method for marketing a B-cell antagonist for use in a rheumatoid arthritis patient subpopulation, the method comprising informing a target audience about the use of the antagonist for treating the patient subpopulation characterized by the presence, in serum samples from patients of such subpopulation, of an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination of these amounts is provided. In a further embodiment, the serum samples from patients of such subpopulation are positive for RF.

In another aspect, a method is provided for selecting a therapy for a patient or a patient subpopulation with rheumatoid arthritis comprising: (a) determining in a serum sample from the patient the amount of CXCL13, sFcRH5, or both of these amounts; (b) determining whether the serum sample is RF positive or RF negative; and (c) selecting a B-cell antagonist as the therapy if the patient's sample is RF positive and has an amount of CXCL13 greater than 116.6 pg/ml, or an amount of sFcRH5 greater than 126.7 ng/ml, or a combination of these amounts in the sample.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a dendrogram depicting sample clusters and branch support values (88, 94, 87, and 67) following microarray analysis of synovial tissues from RA patients as described in Example 1.

FIG. 2 shows a heatmap and bootstrapped dendrogram (vertical lines) revealing four molecular phenotypes (subtypes) of RA as described in Example 1. F1=fibroblast-rich type 1 subtype; F2=fibroblast-rich type 2 subtype; L=lymphoid-rich subtype; M=myeloid-rich subtype. Each molecular phenotype is indicated at the top of the figure above the bootstrapped dendrogram; corresponding boxes around gene expression within the heatmap are indicated and highlight specific areas of coregulated signature genes. Expression data was z-score normalized for visualization (bar at bottom of figure).

FIG. 3 shows molecular, clinical, histological, and immunohistochemical characteristics of L subtype synovial tissue samples as described in Example 1. (A) Expression of XBP1 transcription factor in L subtype samples (L) compared to non-L subtype samples (NL); (B) Expression of XBP1 transcription factor in synovial samples containing lymphoid aggregates (+) compared to synovial samples lacking lymphoid aggregates (−); (C) Graphical plot of erythroid sedimentation rate (ESR) (“Sed Rate”) compared to XBP1 expression level in all RA samples tested; (D) Graphical plot of C-reactive protein compared to XBP1 expression level in all RA samples tested; (E) hematoxylin and eosin staining of a representative synovial sample of the L subtype; (F) immunohistochemical staining for the T cell marker CD3 of a representative synovial sample of the L subtype; (G) immunohistochemical staining for the activated leukocyte marker CD68 of a representative synovial sample of the L subtype; (H) immunohistochemical staining for the B cell marker CD20 of a representative sample of the L subtype.

FIG. 4 shows molecular, histological, and immunohistochemical characteristics of M subtype synovial tissue samples as described in Example 1. (A) Expression of ICAM1 in M subtype samples (M) compared to the expression in the other subtypes (F1, F2, L); (B) Graphical plot of IL1β gene expression compared to TNF gene expression in M subtype samples; (C) hematoxylin and eosin staining of a representative synovial sample of the M subtype; (D) immunohistochemical staining for the T cell marker CD3 of a representative synovial sample of the M subtype; (E) immunohistochemical staining for the activated leukocyte marker CD68 of a representative synovial sample of the M subtype; (F) immunohistochemical staining for the B cell marker CD20 of a representative synovial sample of the M subtype.

FIG. 5 shows molecular, histological, and immunohistochemical characteristics of F2 subtype synovial tissue samples as described in Example 1. (A) Expression of IL17D in F2 subtype samples (F2) compared to the expression in the other subtypes (F1, L, and M); (B) hematoxylin and eosin staining of a representative synovial sample of the F2 subtype; (C) immunohistochemical staining for the T cell marker CD3 of a representative synovial sample of the F2 subtype; (D) immunohistochemical staining for the activated leukocyte marker CD68 of a representative synovial sample of the F2 subtype; (E) immunohistochemical staining for the B cell marker CD20 of a representative synovial sample of the F2 subtype.

FIG. 6 shows molecular, histological, and immunohistochemical characteristics of F1 subtype synovial tissue samples as described in Example 1. (A) Expression of ITGA11 in F1 subtype samples (F1) compared to the expression in the other subtypes (F2, L, and M); (B) hematoxylin and eosin staining of a representative synovial sample of the F1 subtype; (C) immunohistochemical staining for the T cell marker CD3 of a representative synovial sample of the F1 subtype; (D) immunohistochemical staining for the activated leukocyte marker CD68 of a representative synovial sample of the F1 subtype; (E) immunohistochemical staining for the B cell marker CD20 of a representative synovial sample of the F1 subtype.

FIG. 7 shows the percentage of samples with lymphoid clusters according to molecular subtype as described in Example 1. F1, F2, L, and M molecular subtypes are indicated along the bottom axis.

FIG. 8 shows, within each indicated molecular subtype, the value of certain samples for certain classical markers of RA as described in Example 1. (A) erythroid sedimentation rate (ESR) in mm/hr; (B) C-reactive protein (CRP) in mg/dL; (C) radiographic progression by stage. F1, F2, L, and M molecular subtypes are indicated in each of (A)-(C) along the bottom axis; each dot represents the value for one individual sample.

FIG. 9 shows biological pathways within each molecular subtype identified by statistical analysis of gene signatures specific to each subtype as described in Example 1. The heatmap depicts the results of the analysis. Each of the subtypes F1, F2, L, and M is listed across the top of the heatmap; biological pathways are indicated along the right side of the heatmap; grey shading within the heatmap corresponds to the p-values for statistically enriched pathways within each subtype according to the scale shown at the bottom of the figure.

FIG. 10 shows the validation of selected genes found to be differentially expressed by microarray analysis as described in Example 2, (A) F1-specific transcripts; (B) F2-specific transcripts; (C) L-specific transcripts; (D) M-specific transcripts. In each of (A)-(D), the name of the gene transcript is indicated at the top of the graph; each of the subtypes F1, F2, L, and M, along with normal (Nrml) and osteoarthritis (OA) individuals, is indicated along the horizontal axis of each graph; transcript abundance relative to the house keeping gene, HPRT1, is indicated along the vertical axis of each graph.

FIG. 11 shows a graphical plot of (A) serum sFcRH5 levels and (B) serum CXCL13 levels in RA patients in the REFLEX trial prior to dosing with rituximab as compared to healthy controls as described in Example 3. Serum sFcRH5 levels are plotted on the vertical axis in ng/ml in (A); serum CXCL13 levels are plotted on the vertical axis in pg/ml in (B); healthy controls and RA patients are indicated on the horizontal axis in (A) and (B).

FIG. 12 shows the results of the threshold sensitivity analysis of CXCL13 and sFcRH5 data as described in Example 3. Striped bars: rituximab-treated patients; open bars: placebo-treated patients; the width of the bars reflect the number of patients in the group; the right side of the figure shows the placebo-corrected optimal subgroup efficacy difference between the biomarker-high group and the biomarker-low group with 95% confidence intervals (CI).

FIG. 13 shows placebo-controlled 24 week ACR50 response rates in patient subsets defined by sFcRH5 level and RF seropositivity in the REFLEX trial (A) and in the SERENE trial (B) as described in Example 3. Striped bars, rituximab-treated patients; open bars, placebo-treated patients. The patient subsets are indicated along the horizontal axis (all patients, FcRH5 or hi, RF negative or positive); the number of patients in each subset showing an ACR50 response compared to the total number of patients in that subset is indicated above each bar. The placebo-controlled ACR50 response rate (ΔACR50) is also indicated for each subset at the top of the graph.

FIG. 14 shows placebo-controlled 24 week ACR50 response rates in patient subsets defined by sFcRH5 level, CXCL13 level and RF seropositivity in the REFLEX trial as described in Example 3. Striped bars, rituximab-treated patients; open bars, placebo-treated patients. The patient subsets are indicated along the horizontal axis (all patients, FcRH5 lo or hi, CXCL13 lo or hi, RF negative or positive); the number of patients in each subset showing an ACR50 response compared to the total number of patients in that subset is indicated above each bar. The placebo-controlled ACR50 response rate (ΔACR50) is also indicated for each subset at the top of the graph.

DETAILED DESCRIPTION

Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons (New York, N.Y. 1994), and March, Advanced Organic Chemistry Reactions, Mechanisms and Structure 4th ed., John Wiley & Sons (New York, N.Y. 1992), provide one skilled in the art with a general guide to many of the terms used in the present application.

Certain Definitions

For purposes of interpreting this specification, the following definitions will apply and whenever appropriate, terms used in the singular will also include the plural and vice versa. In the event that any definition set forth below conflicts with any document incorporated herein by reference, the definition set forth below shall control.

“Rheumatoid arthritis,” (RA) refers to a chronic systemic autoimmune inflammatory disease that mainly involves the synovial membrane of multiple joints with resultant injury to the articular cartilage, resulting in joint destruction. The main presenting symptoms in RA are pain, stiffness, swelling, and/or loss of function of one or more joints.

The term “polynucleotide” or “nucleic acid,” as used interchangeably herein, refers to polymers of nucleotides of any length, and include DNA and RNA. The nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and/or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase. A polynucleotide may comprise modified nucleotides, such as methylated nucleotides and their analogs. If present, modification to the nucleotide structure may be imparted before or after assembly of the polymer. The sequence of nucleotides may be interrupted by non-nucleotide components. A polynucleotide may be further modified after polymerization, such as by conjugation with a labeling component. Other types of modifications include, for example, “caps”, substitution of one or more of the naturally occurring nucleotides with an analog, internucleotide modifications such as, for example, those with uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoamidates, cabamates, etc.) and with charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), those containing pendant moieties, such as, for example, proteins (e.g., nucleases, toxins, antibodies, signal peptides, poly-L-lysine, etc.), those with intercalators (e.g., acridine, psoralen, etc.), those containing chelators (e.g., metals, radioactive metals, boron, oxidative metals, etc.), those containing alkylators, those with modified linkages (e.g., alpha anomeric nucleic acids, etc.), as well as unmodified forms of the polynucleotide(s). Further, any of the hydroxyl groups ordinarily present in the sugars may be replaced, for example, by phosphonate groups, phosphate groups, protected by standard protecting groups, or activated to prepare additional linkages to additional nucleotides, or may be conjugated to solid supports. The 5′ and 3′ terminal OH can be phosphorylated or substituted with amines or organic capping groups moieties of from 1 to 20 carbon atoms. Other hydroxyls may also be derivatized to standard protecting groups. Polynucleotides can also contain analogous forms of ribose or deoxyribose sugars that are generally known in the art, including, for example, 2′-O-methyl-2′-O— allyl, 2′-fluoro- or 2′-azido-ribose, carbocyclic sugar analogs, α-anomeric sugars, epimeric sugars such as arabinose, xyloses or lyxoses, pyranose sugars, furanose sugars, sedoheptuloses, acyclic analogs and abasic nucleoside analogs such as methyl riboside. One or more phosphodiester linkages may be replaced by alternative linking groups. These alternative linking groups include, but are not limited to, embodiments wherein phosphate is replaced by P(O)S (“thioate”), P(S)S (“dithioate”), “(O)NR 2 (“amidate”), P(O)R, P(O)OR′, CO or CH2 (“formacetal”), in which each R or R′ is independently H or substituted or unsubstituted alkyl (1-20 C) optionally containing an ether (—O—) linkage, aryl, alkenyl, cycloalkyl, cycloalkenyl or araldyl. Not all linkages in a polynucleotide need be identical. The preceding description applies to all polynucleotides referred to herein, including RNA and DNA.

“Oligonucleotide,” as used herein, refers to short, single stranded polynucleotides that are at least about seven nucleotides in length and less than about 250 nucleotides in length. Oligonucleotides may be synthetic. The terms “oligonucleotide” and “polynucleotide” are not mutually exclusive. The description above for polynucleotides is equally and fully applicable to oligonucleotides.

The term “primer” refers to a single stranded polynucleotide that is capable of hybridizing to a nucleic acid and allowing the polymerization of a complementary nucleic acid, generally by providing a free 3′-OH group.

The term “array” or “microarray” refers to an ordered arrangement of hybridizable array elements, preferably polynucleotide probes (e.g., oligonucleotides), on a substrate. The substrate can be a solid substrate, such as a glass slide, or a semi-solid substrate, such as nitrocellulose membrane.

The term “amplification” refers to the process of producing one or more copies of a reference nucleic acid sequence or its complement. Amplification may be linear or exponential (e.g., PCR). A “copy” does not necessarily mean perfect sequence complementarity or identity relative to the template sequence. For example, copies can include nucleotide analogs such as deoxyinosine, intentional sequence alterations (such as sequence alterations introduced through a primer comprising a sequence that is hybridizable, but not fully complementary, to the template), and/or sequence errors that occur during amplification.

The term “detection” includes any means of detecting, including direct and indirect detection.

“Elevated expression” or “elevated levels” refers to an increased expression of a mRNA or a protein in a patient relative to a control, such as an individual or individuals who are not suffering from RA.

The term “molecular subtype,” used interchangeably with “molecular phenotype,” refers to a subtype or phenotype of RA characterized by the expression of one or more particular genes or one or more particular proteins, or a particular pattern of expression of a combination of genes or a combination of proteins. The expression of particular genes, proteins or combinations of genes or proteins may be further associated with certain pathological, histological, and/or clinical features of RA.

The term “multiplex-PCR” refers to a single PCR reaction carried out on nucleic acid obtained from a single source (e.g., a patient) using more than one primer set for the purpose of amplifying two or more DNA sequences in a single reaction.

As used herein, “rheumatoid factor,” or “RF,” refers to IgM, IgG, or IgA isotypes, singly or in any combination, of antibodies detected in patient serum and directed to antigenic determinants present on human and animal IgG.

The term “positive for RF” refers to a result of an assay for RF, e.g., an ELISA assay, where the result is above a threshold or cutoff value for that assay for samples that are considered to reproducibly contain detectable levels of RF.

The term “negative for RF” refers to a result of an assay for RF, e.g., an ELISA assay, where the result is at or below a threshold or cutoff value for that assay for samples that are considered to reproducibly contain undetectable levels of RF.

“Stringency” of hybridization reactions is readily determinable by one of ordinary skill in the art, and generally is an empirical calculation dependent upon probe length, washing temperature, and salt concentration. In general, longer probes require higher temperatures for proper annealing, while shorter probes need lower temperatures. Hybridization generally depends on the ability of denatured DNA to reanneal when complementary strands are present in an environment below their melting temperature. The higher the degree of desired homology between the probe and hybridizable sequence, the higher the relative temperature which can be used. As a result, it follows that higher relative temperatures would tend to make the reaction conditions more stringent, while lower temperatures less so. For additional details and explanation of stringency of hybridization reactions, see Ausubel et al., Current Protocols in Molecular Biology, Wiley Interscience Publishers, (1995).

“Stringent conditions” or “high stringency conditions”, as defined herein, can be identified by those that: (1) employ low ionic strength and high temperature for washing, for example 0.015 M sodium chloride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 50 C; (2) employ during hybridization a denaturing agent, such as formamide, for example, 50% (v/v) formamide with 0.1% bovine serum albumin/0.1% Ficoll/0.1% polyvinylpynolidone/50 mM sodium phosphate buffer at pH 6.5 with 750 mM sodium chloride, 75 mM sodium citrate at 42 C; or (3) overnight hybridization in a solution that employs 50% formamide, 5×SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5×Denhardt's solution, sonicated salmon sperm DNA (50 μg/ml), 0.1% SDS, and 10% dextran sulfate at 42 C, with a 10 minute wash at 42 C in 0.2×SSC (sodium chloride/sodium citrate) followed by a 10 minute high-stringency wash consisting of 0.1×SSC containing EDTA at 55 C.

“Moderately stringent conditions” can be identified as described by Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Press, 1989, and include the use of washing solution and hybridization conditions (e.g., temperature, ionic strength and % SDS) less stringent that those described above. An example of moderately stringent conditions is overnight incubation at 37° C. in a solution comprising: 20% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 mg/ml denatured sheared salmon sperm DNA, followed by washing the filters in 1×SSC at about 37-50 C. The skilled artisan will recognize how to adjust the temperature, ionic strength, etc. as necessary to accommodate factors such as probe length and the like.

The term “biomarker” as used herein refers to an indicator of e.g, a pathological state of a patient, which can be detected in a biological sample of the patient. Biomarkers include, but are not limited to, DNA, RNA, protein, carbohydrate, or glycolipid-based molecular markers.

The term “diagnosis” is used herein to refer to the identification or classification of a molecular or pathological state, disease or condition. For example, “diagnosis” may refer to identification of a particular type of RA. “Diagnosis” may also refer to the classification of a particular subtype of RA, e.g., by histopathological criteria (e.g., lymphoid infiltration or follicle-like lymphoid cluster), or by molecular features (e.g., a subtype characterized by expression of one or a combination of particular genes or proteins encoded by said genes).

The term “aiding diagnosis” is used herein to refer to methods that assist in making a clinical determination regarding the presence, or nature, of a particular type of symptom or condition of RA. For example, a method of aiding diagnosis of RA can comprise measuring the expression of certain genes in a biological sample from an individual.

The term “prognosis” is used herein to refer to the prediction of the likelihood of autoimmune disorder-attributable disease symptoms of an autoimmune disease such as RA. The term “prediction” is used herein to refer to the likelihood that a patient will respond either favorably or unfavorably to a drug or set of drugs. In one embodiment, the prediction relates to the extent of those responses. In one embodiment, the prediction relates to whether and/or the probability that a patient will survive or improve following treatment, for example treatment with a particular therapeutic agent, and for a certain period of time without disease recurrence. The predictive methods of the invention can be used clinically to make treatment decisions by choosing the most appropriate treatment modalities for any particular patient. The predictive methods of the present invention are valuable tools in predicting if a patient is likely to respond favorably to a treatment regimen, such as a given therapeutic regimen, including for example, administration of a given therapeutic agent or combination, surgical intervention, steroid treatment, etc., or whether long-term survival of the patient, following a therapeutic regimen is likely.

As used herein, “treatment” refers to clinical intervention in an attempt to alter the natural course of the individual or cell being treated, and can be performed before or during the course of clinical pathology. Desirable effects of treatment include preventing the occurrence or recurrence of a disease or a condition or symptom thereof, alleviating a condition or symptom of the disease, diminishing any direct or indirect pathological consequences of the disease, decreasing the rate of disease progression, ameliorating or palliating the disease state, and achieving remission or improved prognosis. In some embodiments, methods and compositions of the invention are useful in attempts to delay development of a disease or disorder.

An “effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result. A “therapeutically effective amount” of a therapeutic agent may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the therapeutic agent are outweighed by the therapeutically beneficial effects. A “prophylactically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically but not necessarily, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount.

An “individual,” “subject” or “patient” is a vertebrate. In certain embodiments, the vertebrate is a mammal. Mammals include, but are not limited to, primates (including human and non-human primates) and rodents (e.g., mice and rats). In certain embodiments, a mammal is a human.

A “control subject” refers to a healthy subject who has not been diagnosed as having RA and who does not suffer from any sign or symptom associated with RA.

The term “sample,” as used herein, refers to a composition that is obtained or derived from a subject of interest that contains a cellular and/or other molecular entity that is to be characterized and/or identified, for example based on physical, biochemical, chemical and/or physiological characteristics. For example, the phrase “disease sample” and variations thereof refers to any sample obtained from a subject of interest that would be expected or is known to contain the cellular and/or molecular entity that is to be characterized.

By “tissue” or “cell sample” is meant a collection of similar cells obtained from a tissue of a subject or patient. The source of the tissue or cell sample may be solid tissue as from a fresh, frozen and/or preserved organ or tissue sample or biopsy or aspirate; blood or any blood constituents; bodily fluids such as cerebral spinal fluid, amniotic fluid, peritoneal fluid, or interstitial fluid; cells from any time in gestation or development of the subject. The tissue sample may also be primary or cultured cells or cell lines. Optionally, the tissue or cell sample is obtained from a disease tissue/organ. The tissue sample may contain compounds which are not naturally intermixed with the tissue in nature such as preservatives, anticoagulants, buffers, fixatives, nutrients, antibiotics, or the like. A “reference sample”, “reference cell”, “reference tissue”, “control sample”, “control cell”, or “control tissue”, as used herein, refers to a sample, cell or tissue obtained from a source known, or believed, not to be afflicted with the disease or condition for which a method or composition of the invention is being used to identify. In one embodiment, a reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy part of the body of the same subject or patient in whom a disease or condition is being identified using a composition or method of the invention. In one embodiment, a reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy part of the body of an individual who is not the subject or patient in whom a disease or condition is being identified using a composition or method of the invention.

For the purposes herein a “section” of a tissue sample is meant a single part or piece of a tissue sample, e.g. a thin slice of tissue or cells cut from a tissue sample. It is understood that multiple sections of tissue samples may be taken and subjected to analysis according to the present invention, provided that it is understood that the present invention comprises a method whereby the same section of tissue sample is analyzed at both morphological and molecular levels, or is analyzed with respect to both protein and nucleic acid.

By “correlate” or “correlating” is meant comparing, in any way, the performance and/or results of a first analysis or protocol with the performance and/or results of a second analysis or protocol. For example, one may use the results of a first analysis or protocol in carrying out a second protocols and/or one may use the results of a first analysis or protocol to determine whether a second analysis or protocol should be performed. With respect to the embodiment of gene expression analysis or protocol, one may use the results of the gene expression analysis or protocol to determine whether a specific therapeutic regimen should be performed.

A “medicament” is an active drug to treat a disease, disorder, and/or condition. In one embodiment, the disease, disorder, and/or condition is RA or its symptoms or side effects.

The term “increased resistance” to a particular therapeutic agent or treatment option, when used in accordance with the invention, means decreased response to a standard dose of the drug or to a standard treatment protocol.

The term “decreased sensitivity” to a particular therapeutic agent or treatment option, when used in accordance with the invention, means decreased response to a standard dose of the agent or to a standard treatment protocol, where decreased response can be compensated for (at least partially) by increasing the dose of agent, or the intensity of treatment.

“Patient response” or “response” can be assessed using any endpoint indicating a benefit to the patient, including, without limitation, (1) inhibition, to some extent, of disease progression, including slowing down and complete arrest; (2) reduction in the number of disease episodes and/or symptoms; (3) reduction in lesional size; (4) inhibition (i.e., reduction, slowing down or complete stopping) of disease cell infiltration into adjacent peripheral organs and/or tissues; (5) inhibition (i.e. reduction, slowing down or complete stopping) of disease spread; (6) decrease of auto-immune response, which may, but does not have to, result in the regression or ablation of the disease lesion; (7) relief, to some extent, of one or more symptoms associated with the disorder; (8) increase in the length of disease-free presentation following treatment; and/or (9) decreased mortality at a given point of time following treatment.

The term “gene signature” is used interchangeably with “gene expression signature” and refers to one or a combination of genes whose expression is indicative of a particular subtype of RA characterized by certain molecular, pathological, histological, and/or clinical features. In certain embodiments, the expression of one or more genes comprising the gene signature is elevated compared to that in control subjects.

The term “protein signature” is used interchangeably with “protein expression signature” and refers to one or a combination of proteins whose expression is indicative of a particular subtype of RA characterized by certain molecular, pathological, histological, and/or clinical features. In certain embodiments, the expression of one or more proteins comprising the protein signature is elevated compared to that in control subjects.

A “RA therapeutic agent,” a “therapeutic agent effective to treat RA,” and grammatical variations thereof, as used herein, refer to an agent that when provided in an effective amount is known, clinically shown, or expected by clinicians to provide a therapeutic benefit in a subject who has RA.

A “B-cell surface marker” or “B-cell surface antigen” herein is an antigen expressed on the surface of a B cell that can be targeted with an antagonist that binds thereto. Exemplary B-cell surface markers include the CD10, CD19, CD20 (MS4A1), CD21, CD22, CD23, CD24, CD37, CD40, CD53, CD72, CD73, CD74, CDw75, CDw76, CD77, CDw78, CD79a, CD79b, CD80, CD81, CD82, CD83, CDw84, CMS, and CD86 leukocyte surface markers (for descriptions, see The Leukocyte Antigen Facts Book, 2nd Edition. 1997, ed. Barclay et al. Academic Press, Harcourt Brace & Co., New York). Other B-cell surface markers include RP105, FcRH2, B-cell CR2, CCR6, P2×5, HLA-DOB, CXCR5, FCER2, BR3, Btig, NAG14, SLGC16270, FcRH1, IRTA2, ATWD578, FcRH3, IRTA1, FcRH6, BCMA, and 239287. The B-cell surface marker of particular interest is preferentially expressed on B cells compared to other non-B-cell tissues of a mammal and may be expressed on both precursor B cells and mature B cells.

An “antibody that binds to a B-cell surface marker” is a molecule that, upon binding to a B-cell surface marker, destroys or depletes B cells in a mammal and/or interferes with one or more B-cell functions, e.g. by reducing or preventing a humoral response elicited by the B cell. The antibody in certain instances is able to deplete B cells (i.e, reduce circulating B-cell levels) in a mammal treated therewith. Such depletion may be achieved via various mechanisms such as antibody-dependent cell-mediated cytotoxicity (ADCC) and/or complement-dependent cytotoxicity (CDC), inhibition of B-cell proliferation, and/or induction of B-cell death (e.g. via apoptosis).

An “antagonist” refers to a molecule capable of neutralizing, blocking, inhibiting, abrogating, reducing or interfering with the activities of a particular or specified protein, including its binding to one or more receptors in the case of a ligand or binding to one or more ligands in case of a receptor. Antagonists include antibodies and antigen-binding fragments thereof, proteins, peptides, glycoproteins, glycopeptides, glycolipids, polysaccharides, oligosaccharides, nucleic acids, bioorganic molecules, peptidomimetics, pharmacological agents and their metabolites, transcriptional and translation control sequences, and the like. Antagonists also include small molecule inhibitors of the protein, and fusion proteins, receptor molecules and derivatives which bind specifically to the protein thereby sequestering its binding to its target, antagonist variants of the protein, antisense molecules directed to the protein, RNA aptamers, and ribozymes against the protein.

A “B-cell antagonist” is a molecule that, upon binding to a B-cell surface marker, destroys or depletes B cells in a mammal and/or interferes with one or more B-cell functions, e.g. by reducing or preventing a humoral response elicited by the B cell. The antagonist in certain instances is able to deplete B cells (i.e. reduce circulating B-cell levels) in a mammal treated therewith. Such depletion may be achieved via various mechanisms such as ADCC and/or CDC, inhibition of B-cell proliferation, and/or induction of B-cell death (e.g. via apoptosis). Exemplary antagonists include synthetic or native-sequence peptides, fusion proteins, and small-molecule antagonists that bind to the B-cell marker, optionally conjugated with or fused to a cytotoxic agent. Examples include but are not limited to, e.g., CD22 antibodies, CD20 antibodies, BR3 antibodies (e.g., WO0224909), and BR3-Fc immunoadhesin.

Examples of CD20 antibodies include: “C2B8,” which is now called “rituximab” (“RITUXAN®”) (U.S. Pat. No. 5,736,137); the yttrium-[90]-labeled 2B8 murine antibody designated “Y2B8” or “ibritumomab tiuxetan” (ZEVALIN®) commercially available from IDEC Pharmaceuticals, Inc. (U.S. Pat. No. 5,736,137; 2B8 deposited with ATCC under accession no. HB11388 on Jun. 22, 1993); murine IgG2a “B1,” also called “tositumomab,” optionally labeled with ¹³¹I to generate the “¹³I—B1” or “iodine I¹³¹ tositumomab” antibody (BEXXAR™) commercially available from Corixa (see, also, U.S. Pat. No. 5,595,721); murine monoclonal antibody “1F5” (Press et al. Blood 69(2):584-591 (1987) and variants thereof including “framework-patched” or humanized 1F5 (WO 2003/002607, Leung, S.; ATCC deposit HB-96450); murine 2H7 and chimeric 2H7 antibody (U.S. Pat. No. 5,677,180); humanized 2H7 (see, e.g., WO04/056312; US20060024295); HUMAX-CD20™ antibodies (Genmab, Denmark); the human monoclonal antibodies set forth in WO 2004/035607 (Teeling et al.); AME-133™ antibodies (Applied Molecular Evolution); A20 antibody or variants thereof such as chimeric or humanized A20 antibody (cA20, hA20, respectively) (US 2003/0219433, Immunomedics); and monoclonal antibodies L27, G28-2, 93-1 B3, B-C1 or NU-B2 available from the International Leukocyte Typing Workshop (Valentine et al., In: Leukocyte Typing III (McMichael, Ed., p. 440, Oxford University Press (1987)).

The terms “BAFF,” “BAFF polypeptide,” “TALL-1” or “TALL-1 polypeptide,” “BLyS”, and “THANK” when used herein encompass “native-sequence BAFF polypeptides” and “BAFF variants.” “BAFF” is a designation given to those polypeptides that have the human BAFF sequence as set forth in, for example, U.S. Pat. Pub. No. 2006/0110387, and homologs and fragments and variants thereof, which have the biological activity of the native-sequence BAFF. A biological activity of BAFF can be selected from the group consisting of promoting B-cell survival, promoting B-cell maturation, and binding to BR3. The term “BAFF” includes those polypeptides described in Shu et al., J. Leukocyte Biol., 65:680 (1999); GenBank Accession No. AF136293; WO 1998/18921; EP 869,180; WO 1998/27114; WO 1999/12964; WO 1999/33980; Moore et al., Science, 285:260-263 (1999); Schneider et al., J. Exp. Med., 189:1747-1756 (1999); and Mukhopadhyay et al., J. Biol. Chem., 274:15978-15981 (1999).

The term “BAFF antagonist” as used herein is used in the broadest sense, and includes any molecule that (1) binds a native-sequence BAFF polypeptide or binds a native-sequence BR3 polypeptide to block, partially or fully, BR3 interaction with BAFF polypeptide, and (2) partially or fully blocks, inhibits, or neutralizes native-sequence BAFF signaling. Native-sequence BAFF polypeptide signaling promotes, among other things, B-cell survival and B-cell maturation. The inhibition, blockage, or neutralization of BAFF signaling results in, inter alia, a reduction in the number of B cells. A BAFF antagonist as defined herein will partially or fully block, inhibit, or neutralize one or more biological activities of a BAFF polypeptide, in vitro or in vivo. In one embodiment, a biologically active BAFF potentiates any one or a combination of the following events in vitro or in vivo: an increased survival of B cells, an increased level of IgG and/or IgM, an increased numbers of plasma cells, and processing of NF-κb2/100 to p52 NF-κβ in splenic B cells (e.g., Batten et al., J. Exp. Med. 192:1453-1465 (2000); Moore et al., Science 285:260-263 (1999); and Kayagaki et al., Immunity, 10:515-524 (2002)).

In some embodiments, a BAFF antagonist as defined herein includes anti-BAFF antibodies, BAFF-binding polypeptides (including immunoadhesins and peptides), and BAFF-binding small molecules. BAFF antagonists include, for example, the BAFF-binding antibodies described in WO 2002/02641 (e.g., antibodies comprising the amino acid sequence of any of SEQ ID NOS:1-46, 321-329, 834-872, 1563-1595, 1881-1905 of Table 1 thereof). In a further embodiment, the immunoadhesin comprises a BAFF-binding region of a BAFF receptor (e.g., an extracellular domain of BR3, BCMA, or TACI). In a still further embodiment, the immunoadhesin is BR3-Fc. Other examples of BAFF-binding Fc proteins can be found in WO 2002/66516, WO 2000/40716, WO 2001/87979, WO 2003/024991, WO 2002/16412, WO 2002/38766, WO 2002/092620, and WO 2001/12812. Methods of making BAFF antagonists are described, for example, in US 2005/0095243 and US 2005/0163775.

The terms “BR3”, “BR3 polypeptide” or “BR3 receptor” when used herein encompass native-sequence BR3 polypeptides and BR3 variants, as defined hereinbelow. “BR3” is a designation given to those polypeptides comprising, for example, the human BR3 sequence set forth in WO 2003/14294 and US 2005/0070689. BR3 polypeptides can be isolated from a variety of sources, such as from human tissue types or from another source, or prepared by recombinant and/or synthetic methods. The term BR3 includes the BR3 polypeptides described in WO 2002/24909, WO 2003/14294, and US 2005/0070689. Anti-BR3 antibodies can be prepared in accordance with methods set for in, for example, WO 2003/14294 and US 2005/0070689.

A “native-sequence” BR3 polypeptide or “native BR3” comprises a polypeptide having the same amino acid sequence as the corresponding BR3 polypeptide derived from nature. Such native-sequence BR3 polypeptides can be isolated from nature or can be produced by recombinant and/or synthetic means. The term “native-sequence BR3 polypeptide” specifically encompasses naturally occurring truncated, soluble or secreted forms (e.g., an extracellular domain sequence), naturally occurring variant forms (e.g., alternatively spliced forms) and naturally occurring allelic variants of the polypeptide. The BR3 polypeptides of the invention include the BR3 polypeptide comprising or consisting of the contiguous sequence of amino acid residues 1 to 184 of a human BR3 (see WO 2003/14294 and US 2005/0070689).

A BR3 “extracellular domain” or “ECD” refers to a form of the BR3 polypeptide that is essentially free of the transmembrane and cytoplasmic domains. ECD forms of BR3 include a polypeptide comprising any one of the amino acid sequences selected from the group consisting of amino acids 1-77, 2-62, 2-71, 1-61, 7-71, 23-38 and 2-63 of human BR3. In certain embodiments, BAFF antagonists are polypeptides comprising any one of the above-mentioned ECD forms of human BR3 and variants and fragments thereof that bind a native BAFF.

“BR3 variant” means a BR3 polypeptide having at least about 80% amino acid sequence identity with the amino acid sequence of a native-sequence, full-length BR3 or BR3 ECD and binds a native-sequence BAFF polypeptide. Optionally, the BR3 variant includes a single cysteine-rich domain. Such BR3 variant polypeptides include, for instance, BR3 polypeptides wherein one or more amino acid residues are added, or deleted, at the N- and/or C-terminus, as well as within one or more internal domains, of the full-length amino acid sequence. Fragments of the BR3ECD that bind a native sequence BAFF polypeptide are also contemplated.

The term “APRIL antagonist” as used herein is used in the broadest sense, and includes any molecule that (1) binds a native-sequence APRIL polypeptide or binds a native-sequence ligand to APRIL to block, partially or fully, the ligand's interaction with APRIL polypeptide, and (2) partially or fully blocks, inhibits, or neutralizes native-sequence APRIL signaling. Native-sequence APRIL polypeptide signaling promotes, among other things, B-cell survival and B-cell maturation. APRIL (a proliferation-inducing ligand) is a TNF family member with a shared receptor to BAFF. Examples of APRIL antagonists include but are not limited to atacicept (same as TACI-Ig immunoadhesin) and a BAFF/APRIL antagonist (soluble BCMA-Fc).

The term “cytokine” is a generic term for proteins released by one cell population that act on another cell as intercellular mediators. Examples of such cytokines are lymphokines, monokines; interleukins (ILs) such as IL-1, IL-1a, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12, IL-15, IL-17A, IL-17F, IL-17A/F; a tumor necrosis factor such as TNF-α or TNF-β; and other polypeptide factors including LIF and kit ligand (KL). As used herein, the term cytokine includes proteins from natural sources or from recombinant cell culture and biologically active equivalents of the native-sequence cytokines, including synthetically produced small-molecule entities and pharmaceutically acceptable derivatives and salts thereof.

For the purposes herein, “tumor necrosis factor-alpha (TNF-alpha)” refers to a human TNF-alpha molecule comprising the amino acid sequence as described in Pennica et al., Nature, 312:724 (1984) or Aggarwal et al., JBC, 260:2345 (1985).

A “TNF-alpha inhibitor” herein is an agent that inhibits, to some extent, a biological function of TNF-alpha, generally through binding to TNF-alpha and neutralizing its activity. Examples of TNF inhibitors specifically contemplated herein are etanercept (ENBREL®), infliximab (REMICADE®), adalimumab (HUMIRA®), golimumab (SIMPONI™), and certolizumab pegol)(CIMZIA®).

An “IL-17A/F binding agent” is an agent, e.g., an antibody, that binds to the cytokine IL-17A/F or an agent that is cross-reactive with TL-17A and IL-17F.

An “IL-6 binding agent” is an agent, e.g., an antibody, that binds to the cytokine IL-6.

A “CD4 binding agent” is an agent, e.g., an antibody, that binds to the surface glycoprotein CD4 expressed on cells of the T lymphocyte lineage.

Examples of “disease-modifying anti-rheumatic drugs” or “DMARDs” include hydroxycloroquine, sulfasalazine, methotrexate (plus oral and subcutaneous methrotrexate), leflunomide, azathioprine, D-penicillamine, Gold (oral), Gold (intramuscular), minocycline, cyclosporine, Staphylococcal protein A immunoadsorption, including salts and derivatives thereof, etc.

“CTLA4” is expressed on activated T lymphocytes and is involved in down-regulation of the immune response. Other names for CTLA4 in the literature include cytotoxic T-lymphocyte-associated antigen 4, cytotoxic T-lymphocyte-associated protein 4, cell differentiation antigen CD152, and cytotoxic T-lymphocyte-associated granule serine protease 4.

A therapeutic agent that has “marketing approval,” or that has been “approved as a therapeutic agent,” or grammatical variations thereof of these phrases, as used herein, refer to an agent (e.g., in the form of a drug formulation, medicament) that is approved, licensed, registered or authorized by a relevant governmental entity (e.g., federal, state or local regulatory agency, department, bureau) to be sold by and/or through and/or on behalf of a commercial entity (e.g., a for-profit entity) for the treatment of a particular disorder (e.g., RA) or a patient subpopulation (e.g., patients of a particular ethnicity, gender, lifestyle, disease risk profile, etc.). A relevant governmental entity includes, for example, the Food and Drug Administration (FDA), European Medicines Evaluation Agency (EMEA), and equivalents thereof.

“Antibodies” (Abs) and “immunoglobulins” (Igs) refer to glycoproteins having similar structural characteristics. While antibodies exhibit binding specificity to a specific antigen, immunoglobulins include both antibodies and other antibody-like molecules which generally lack antigen specificity. Polypeptides of the latter kind are, for example, produced at low levels by the lymph system and at increased levels by myelomas.

The terms “antibody” and “immunoglobulin” are used interchangeably in the broadest sense and include monoclonal antibodies (e.g., full length or intact monoclonal antibodies), polyclonal antibodies, monovalent antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific antibodies so long as they exhibit the desired biological activity) and may also include certain antibody fragments (as described in greater detail herein). An antibody can be chimeric, human, humanized and/or affinity matured.

The terms “full length antibody,” “intact antibody” and “whole antibody” are used herein interchangeably to refer to an antibody in its substantially intact form, not antibody fragments as defined below. The terms particularly refer to an antibody with heavy chains that contain the Fc region.

“Antibody fragments” comprise a portion of an intact antibody, preferably comprising the antigen binding region thereof. Examples of antibody fragments include Fab, Fab′, F(ab′)₂, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

Papain digestion of antibodies produces two identical antigen-binding fragments, called “Fab” fragments, each with a single antigen-binding site, and a residual “Fc” fragment, whose name reflects its ability to crystallize readily. Pepsin treatment yields an F(ab′)₂ fragment that has two antigen-combining sites and is still capable of cross-linking antigen.

“Fv” is a minimum antibody fragment which contains a complete antigen-binding site. In one embodiment, a two-chain Fv species consists of a dimer of one heavy- and one light-chain variable domain in tight, non-covalent association. Collectively, the six CDRs of an Fv confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

The Fab fragment contains the heavy- and light-chain variable domains and also contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab′ fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CH1 domain including one or more cysteines from the antibody hinge region. Fab′-SH is the designation herein for Fab′ in which the cysteine residue(s) of the constant domains bear a free thiol group. F(ab′)₂ antibody fragments originally were produced as pairs of Fab′ fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible mutations, e.g., naturally occurring mutations, that may be present in minor amounts. Thus, the modifier “monoclonal” indicates the character of the antibody as not being a mixture of discrete antibodies. In certain embodiments, such a monoclonal antibody typically includes an antibody comprising a polypeptide sequence that binds a target, wherein the target-binding polypeptide sequence was obtained by a process that includes the selection of a single target binding polypeptide sequence from a plurality of polypeptide sequences. For example, the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, or recombinant DNA clones. It should be understood that a selected target binding sequence can be further altered, for example, to improve affinity for the target, to humanize the target binding sequence, to improve its production in cell culture, to reduce its immunogenicity in vivo, to create a multispecific antibody, etc., and that an antibody comprising the altered target binding sequence is also a monoclonal antibody of this invention. In contrast to polyclonal antibody preparations which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibody preparations are advantageous in that they are typically uncontaminated by other immunoglobulins.

The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by a variety of techniques, including, for example, the hybridoma method (e.g., Kohler et al., Nature, 256: 495 (1975); Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2^(nd) ed. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981)), recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567), phage display technologies (see, e.g., Clackson et al., Nature, 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132 (2004), and technologies for producing human or human-like antibodies in animals that have parts or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (see, e.g., WO98/24893; WO96/34096; WO96/33735; WO91/10741; Jakobovits et al., Proc. Natl. Acad. Sci. USA 90: 2551 (1993); Jakobovits et al., Nature 362: 255-258 (1993); Bruggemann et al., Year in Immunol. 7:33 (1993); U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016; Marks et al., Bio. Technology 10: 779-783 (1992); Lonberg et al., Nature 368: 856-859 (1994); Morrison, Nature 368: 812-813 (1994); Fishwild et al., Nature Biotechnol. 14: 845-851 (1996); Neuberger, Nature Biotechnol. 14: 826 (1996) and Lonberg and Huszar, Intern. Rev. Immunol. 13: 65-93 (1995).

The monoclonal antibodies herein specifically include “chimeric” antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA 81:6855-9855 (1984)).

“Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. In one embodiment, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit, or nonhuman primate having the desired specificity, affinity, and/or capacity. In some instances, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications may be made to further refine antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin, and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). See also the following review articles and references cited therein: Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1:105-115 (1998); Harris, Biochem. Soc. Transactions 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994).

A “human antibody” is one which comprises an amino acid sequence corresponding to that of an antibody produced by a human and/or has been made using any of the techniques for making human antibodies as disclosed herein. Such techniques include screening human-derived combinatorial libraries, such as phage display libraries (see, e.g., Marks et al., J. Mol. Biol., 222: 581-597 (1991) and Hoogenboom et al., Nucl. Acids Res., 19: 4133-4137 (1991)); using human myeloma and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies (see, e.g., Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 55-93 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991)); and generating monoclonal antibodies in transgenic animals (e.g., mice) that are capable of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production (see, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90: 2551 (1993); Jakobovits et al., Nature, 362: 255 (1993); Bruggermann et al., Year in Immunol., 7: 33 (1993)). This definition of a human antibody specifically excludes a humanized antibody comprising antigen-binding residues from a non-human animal.

An “affinity matured” antibody is one with one or more alterations in one or more CDRs thereof which result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody which does not possess those alteration(s). In one embodiment, an affinity matured antibody has nanomolar or even picomolar affinities for the target antigen. Affinity matured antibodies are produced by procedures known in the art. Marks et al. Bio/Technology 10:779-783 (1992) describes affinity maturation by VH and VL domain shuffling. Random mutagenesis of HVR and/or framework residues is described by: Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al. Gene 169:147-155 (1995); Yelton et al. J. Immunol. 155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995); and Hawkins et al, J. Mol. Biol. 226:889-896 (1992).

A “blocking antibody” or an “antagonist antibody” is one which inhibits or reduces a biological activity of the antigen it binds. Certain blocking antibodies or antagonist antibodies partially or completely inhibit the biological activity of the antigen.

As used herein, “growth-inhibitory” antibodies are those that prevent or reduce proliferation of a cell expressing an antigen to which the antibody binds. For example, the antibody may prevent or reduce proliferation of B cells in vitro and/or in vivo.

Antibodies that “induce apoptosis” refer to antibodies that induce programmed cell death, e.g. of a B cell, as determined by standard apoptosis assays, such as binding of annexin V, fragmentation of DNA, cell shrinkage, dilation of endoplasmic reticulum, cell fragmentation, and/or formation of membrane vesicles (called apoptotic bodies).

Antibody “effector functions” refer to those biological activities attributable to the Fc region (a native-sequence Fc region or amino-acid-sequence-variant Fc region) of an antibody, and vary with the antibody isotype. Examples of antibody effector functions include but are not limited to: C1q binding and complement-dependent cytotoxicity (CDC); Fc-receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down-regulation of cell-surface receptors (e.g. B-cell receptor); and B-cell activation.

The term “Fc region” herein is used to define a C-terminal region of an immunoglobulin heavy chain, including native-sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy-chain Fc region is typically defined to stretch from an amino acid residue at position Cys226, or from Pro230, to the carboxyl-terminus thereof. The C-terminal lysine (residue 447 according to the EU numbering system) of the Fc region may be removed, for example, during production or purification of the antibody, or by recombinantly engineering the nucleic acid encoding a heavy chain of the antibody. Accordingly, a composition of intact antibodies may comprise antibody populations with all K447 residues removed, antibody populations with no K447 residues removed, and antibody populations having a mixture of antibodies with and without the K447 residue.

Unless indicated otherwise herein, the numbering of the residues in an immunoglobulin heavy chain is that of the EU index as in Kabat (Kabat et al., Sequences of Proteins of Immunological Interest, Ed. 5 (Public Health Service, National Institutes of Health, Bethesda, Md., 1991)). The “EU index as in Kabat” refers to the residue numbering of the human IgG1 EU antibody.

A “functional Fc region” possesses an “effector function” of a native-sequence Fc region. Exemplary “effector functions” include but are not limited to C1q binding; CDC; Fc-receptor binding; ADCC; phagocytosis; down-regulation of cell-surface receptors (e.g. B-cell receptor; BCR), etc. Such effector functions generally require the Fc region to be combined with a binding domain (e.g. an antibody-variable domain) and can be assessed using various assays as disclosed, for example, herein.

A “native-sequence Fc region” comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. Native-sequence human Fc regions include a native-sequence human IgG1 Fc region (non-A and A allotypes); native-sequence human IgG2 Fc region; native-sequence human IgG3 Fc region; and native-sequence human IgG4 Fc region, as well as naturally occurring variants thereof.

A “variant Fc region” comprises an amino acid sequence which differs from that of a native-sequence Fc region by virtue of at least one amino acid modification, typically one or more amino acid substitution(s).

The term “Fe-region-comprising antibody” refers to an antibody that comprises an Fc region. The C-terminal lysine (residue 447 according to the EU numbering system) of the Fc region may be removed, for example, during purification of the antibody or by recombinant engineering the nucleic acid encoding the antibody. Accordingly, a composition comprising an antibody having an Fc region can comprise an antibody with K447, with all K447 removed, or a mixture of antibodies with and without the K447 residue.

“Fc receptor” or “FcR” describes a receptor that binds to the Fc region of an antibody. In some embodiments, an FcR is a native-human FcR. In some embodiments, an FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced forms of those receptors. FcγRII receptors include FcγRIIA (an “activating receptor”) and FcγRIII (an “inhibiting receptor”), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof. Activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. Inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain. (see, e.g., Daëron, Anna. Rev. Immunol. 15:203-234 (1997)). FcRs are reviewed, for example, in Ravetch and Kinet, Anna. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term “FcR” herein.

The term “Fc receptor” or “FcR” also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)) and regulation of homeostasis of immunoglobulins. Methods of measuring binding to FcRn are known (see, e.g., Ghetie and Ward, Immunology Today, 18 (12):592-8 (1997); Ghetie et al., Nature Biotechnology, 15 (7):637-40 (1997); Hinton et al., J. Biol. Chem., 279(8):6213-6 (2004); WO 2004/92219 (Hinton et al.).

Binding to human FcRn in vivo and serum half-life of human FcRn high-affinity binding polypeptides can be assayed, e.g., in transgenic mice or transfected human cell lines expressing human FcRn, or in primates to which the polypeptides with a variant Fc region are administered. WO 2000/42072 (Presta) describes antibody variants with improved or diminished binding to FcRs. See, also, for example, Shields et al., J. Biol. Chem., 9(2): 6591-6604 (2001).

“Human effector cells” are leukocytes which express one or more FcRs and perform effector functions. In certain embodiments, the cells express at least FcγRIII and perform ADCC effector function(s). Examples of human leukocytes which mediate ADCC include peripheral blood mononuclear cells (PBMC), natural-killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils. The effector cells may be isolated from a native source, e.g., from blood.

“Antibody-dependent cell-mediated cytotoxicity” or “ADCC” refers to a form of cytotoxicity in which secreted Ig bound onto Fc receptors (FcRs) present on certain cytotoxic cells (e.g., NK cells, neutrophils, and macrophages) enables these cytotoxic effector cells to bind specifically to an antigen-bearing target cell and subsequently kill the target cell with cytotoxins. The primary cells for mediating ADCC, NK cells, express FcγRIII only, whereas monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol., 9:457-492 (1991). To assess ADCC activity of a molecule of interest, an in vitro ADCC assay, such as that described in U.S. Pat. No. 5,500,362 or 5,821,337 or 6,737,056 (Presta), may be performed. Useful effector cells for such assays include PBMC and NK cells. Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al., Proc. Natl. Acad. Sci. (USA), 95:652-656 (1998).

“Complement-dependent cytotoxicity” or “CDC” refers to the lysis of a target cell in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to antibodies (of the appropriate subclass), which are bound to their cognate antigen. To assess complement activation, a CDC assay, e.g. as described in Gazzano-Santoro et al., J. Immunol. Methods, 202:163 (1996), may be performed. Polypeptide variants with altered Fc region amino acid sequences (polypeptides with a variant Fc region) and increased or decreased C1q binding capability are described, e.g., in U.S. Pat. No. 6,194,551 and WO 1999/51642. See, also, e.g., Idusogie et al., J. Immunol. 164:4178-4184 (2000).

“Binding affinity” generally refers to the strength of the sum total of noncovalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless indicated otherwise, as used herein, “binding affinity” refers to intrinsic binding affinity which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally bind antigen slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind antigen faster and tend to remain bound longer. A variety of methods of measuring binding affinity are known in the art.

The term “substantially similar” or “substantially the same,” as used herein, denotes a sufficiently high degree of similarity between two numeric values (for example, one associated with an antibody of the invention and the other associated with a reference/comparator antibody), such that one of skill in the art would consider the difference between the two values to be of little or no biological and/or statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values). The difference between said two values is, for example, less than about 50%, less than about 40%, less than about 30%, less than about 20%, and/or less than about 10% as a function of the reference/comparator value.

The phrase “substantially reduced,” or “substantially different,” as used herein, denotes a sufficiently high degree of difference between two numeric values (generally one associated with a molecule and the other associated with a reference/comparator molecule) such that one of skill in the art would consider the difference between the two values to be of statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values). The difference between said two values is, for example, greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, and/or greater than about 50% as a function of the value for the reference/comparator molecule.

A “small molecule” or “small organic molecule” is defined herein as an organic molecule having a molecular weight below about 500 Daltons.

The word “label” when used herein refers to a detectable compound or composition. The label is typically conjugated or fused directly or indirectly to a reagent, such as a nucleic acid probe or an antibody, and facilitates detection of the reagent to which it is conjugated or fused. The label may itself be detectable (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which results in a detectable product.

An “isolated” biological molecule, such as a nucleic acid, polypeptide, or antibody, is one which has been identified and separated and/or recovered from at least one component of its natural environment.

Reference to “about” a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se. For example, description referring to “about X” includes description of “X.”

The term “pharmaceutical formulation” refers to a sterile preparation that is in such form as to permit the biological activity of the medicament to be effective, and which contains no additional components that are unacceptably toxic to a subject to which the formulation would be administered.

A “sterile” formulation is aseptic or free from all living microorganisms and their spores.

A “package insert” is used to refer to instructions customarily included in commercial packages of therapeutic products or medicaments, that contain information about the indications, usage, dosage, administration, contraindications, other therapeutic products to be combined with the packaged product, and/or warnings concerning the use of such therapeutic products or medicaments and the like.

A “kit” is any manufacture (e.g a package or container) comprising at least one reagent, e.g., a medicament for treatment of RA or joint damage, or a probe for specifically detecting a biomarker gene or protein of the invention. In certain embodiments, the manufacture is promoted, distributed, or sold as a unit for performing the methods of the present invention.

A “target audience” is a group of people or an institution to whom or to which a particular medicament is being promoted or intended to be promoted, as by marketing or advertising, especially for particular uses, treatments, or indications, such as individual patients, patient populations, readers of newspapers, medical literature, and magazines, television or internet viewers, radio or internet listeners, physicians, drug companies, etc.

The term “serum sample” refers to any serum sample obtained from an individual. Methods for obtaining sera from mammals are well known in the art.

The expression “not responsive to,” as it relates to the reaction of subjects or patients to one or more of the medicaments that were previously administered to them, describes those subjects or patients who, upon administration of such medicament(s), did not exhibit any or adequate signs of treatment of the disorder for which they were being treated, or they exhibited a clinically unacceptably high degree of toxicity to the medicament(s), or they did not maintain the signs of treatment after first being administered such medicament(s), with the word treatment being used in this context as defined herein. The phrase “not responsive” includes a description of those subjects who are resistant and/or refractory to the previously administered medication(s), and includes the situations in which a subject or patient has progressed while receiving the medicament(s) that he or she is being given, and in which a subject or patient has progressed within 12 months (for example, within six months) after completing a regimen involving the medicament(s) to which he or she is no longer responsive. The non-responsiveness to one or more medicaments thus includes subjects who continue to have active disease following previous or current treatment therewith. For instance, a patient may have active disease activity after about one to three months of therapy with the medicament(s) to which they are non-responsive. Such responsiveness may be assessed by a clinician skilled in treating the disorder in question.

For purposes of non-response to medicament(s), a subject who experiences “a clinically unacceptably high level of toxicity” from previous or current treatment with one or more medicaments experiences one or more negative side-effects or adverse events associated therewith that are considered by an experienced clinician to be significant, such as, for example, serious infections, congestive heart failure, demyelination (leading to multiple sclerosis), significant hypersensitivity, neuropathological events, high degrees of autoimmunity, a cancer such as endometrial cancer, non-Hodgkin's lymphoma, breast cancer, prostate cancer, lung cancer, ovarian cancer, or melanoma, tuberculosis (TB), and the like.

By “reducing the risk of a negative side effect” is meant reducing the risk of a side effect resulting from treatment with the antagonist herein to a lower extent than the risk observed resulting from treatment of the same patient or another patient with a previously administered medicament. Such side effects include those set forth above regarding toxicity, and are preferably infection, cancer, heart failure, or demyelination.

The “amount” or “level” of a biomarker associated with an increased clinical benefit to a RA patient or patient with joint damage is a detectable level in a biological sample. These can be measured by methods known to one skilled in the art and also disclosed herein. The expression level or amount of biomarker assessed can be used to determine the response to the treatment.

The terms “level of expression” or “expression level” in general are used interchangeably and generally refer to the amount of a polynucleotide or an amino acid product or protein in a biological sample. “Expression” generally refers to the process by which gene-encoded information is converted into the structures present and operating in the cell. Therefore, as used herein, “expression” of a gene may refer to transcription into a polynucleotide, translation into a protein, or even posttranslational modification of the protein. Fragments of the transcribed polynucleotide, the translated protein, or the post-translationally modified protein shall also be regarded as expressed whether they originate from a transcript generated by alternative splicing or a degraded transcript, or from a post-translational processing of the protein, e.g., by proteolysis. “Expressed genes” include those that are transcribed into a polynucleotide as mRNA and then translated into a protein, and also those that are transcribed into RNA but not translated into a protein (for example, transfer and ribosomal RNAs).

Rheumatoid Arthritis

Autoimmune diseases remain clinically important diseases in humans. As the name implies, autoimmune diseases act through the body's own immune system. While the pathological mechanisms differ among individual types of autoimmune diseases, one general mechanism involves the generation of antibodies (referred to herein as self-reactive antibodies or autoantibodies) directed against specific endogenous proteins. Physicians and scientists have identified more than 70 clinically distinct autoimmune diseases, including RA, multiple sclerosis (MS), vasculitis, immune-mediated diabetes, and lupus such as systemic lupus erythematosus (SLE). While many autoimmune diseases are rare—affecting fewer than 200,000 individuals—collectively, these diseases afflict millions of Americans, an estimated five percent of the population, with women disproportionately affected by most diseases. The chronic nature of these diseases leads to an immense social and financial burden.

Inflammatory arthritis is a prominent clinical manifestation in diverse autoimmune disorders including RA, psoriatic arthritis (PsA), SLE, Sjögren's syndrome, and polymyositis. Most of these patients develop joint deformities on physical examination but typically only RA and PsA patients manifest bone erosions on imaging studies.

RA is a chronic inflammatory disease that affects approximately 0.5 to 1% of the adult population in northern Europe and North America, and a slightly lower proportion in other parts of the world. Alamanos and Drosos, Autoimmun. Rev., 4: 130-136 (2005). It is a systemic inflammatory disease characterized by chronic inflammation in the synovial membrane of affected joints, which ultimately leads to loss of daily function due to chronic pain and fatigue. The majority of patients also experience progressive deterioration of cartilage and bone in the affected joints, which may eventually lead to permanent disability. The long-term prognosis of RA is poor, with approximately 50% of patients experiencing significant functional disability within 10 years from the time of diagnosis. Keystone, Rheumatology, 44 (Suppl. 2): ii8-ii12 (2005). Life expectancy is reduced by an average of 3-10 years. Alamanos and Drosos, supra. Patients with a high titer of rheumatoid factor (RF) (approximately 80% of patients) have more aggressive disease (Bukhari et al., Arthritis Rheum., 46: 906-912 (2002)), with a worse long-term outcome and increased mortality over those who are RF negative. Heliovaara et al., Ann. Rheum. Dis., 54: 811-814 (1995)).

The pathogenesis of chronic inflammatory bone diseases, such as RA, is not fully elucidated. Such diseases are accompanied by bone loss around affected joints due to increased osteoclastic resorption. This process is mediated largely by increased local production of pro-inflammatory cytokines. Teitelbaum, Science, 289:1504-1508 (2000); Goldring and Gravallese, Arthritis Res., 2(1):33-37 (2000). These cytokines can act directly on cells in the osteoclast lineage or indirectly by affecting the production of the essential osteoclast differentiation factor, receptor activator of NFκB ligand (RANKL), and/or its soluble decoy receptor, osteoprotegerin (OPG), by osteoblast/stromal cells. Hofbauer et al., J. Bone Min. Res., 15(1):2-12 (2000). Tumor necrosis factor-alpha (TNF-α) is a major mediator of inflammation. Its importance in the pathogenesis of various forms of bone loss is supported by several lines of experimental and clinical evidence. Feldmann et al., Cell, 85(3):307-310 (1996). However, TNF-α is not essential for osteoclastogenesis (Douni et al., J. Inflamm., 47:27-38 (1996)), erosive arthritis (Campbell et al., J. Clin. Invest., 107(12):1519-1527 (2001)), or osteolysis (Childs et al., J. Bon. Min. Res., 16:338-347 (2001)), as these can occur in the absence of TNF-α.

In RA specifically, an immune response is thought to be initiated/perpetuated by one or several antigens presenting in the synovial compartment, producing an influx of acute inflammatory cells and lymphocytes into the joint. Successive waves of inflammation lead to the formation of an invasive and erosive tissue called pannus. This contains proliferating fibroblast-like synoviocytes and macrophages that produce proinflammatory cytokines such as TNF-α and interleukin-1 (IL-1). Local release of proteolytic enzymes, various inflammatory mediators, and osteoclast activation contributes to much of the tissue damage. There is loss of articular cartilage and the formation of bony erosions. Surrounding tendons and bursa may become affected by the inflammatory process. Ultimately, the integrity of the joint structure is compromised, producing disability.

The precise contributions of B cells to the immunopathogenesis of RA are not completely characterized. However, there are several possible mechanisms by which B cells may participate in the disease process. Silverman and Carson, Arthritis Res. Ther., 5 Suppl. 4: S1-6 (2003).

Historically, B cells were thought to contribute to the disease process in RA predominantly by serving as the precursors of autoantibody-producing cells. A number of autoantibody specificities have been identified including antibodies to Type II collagen, and proteoglycans, as well as RFs. The generation of large quantities of antibody leads to immune complex formation and the activation of the complement cascade. This in turn amplifies the immune response and may culminate in local cell lysis. Increased RF synthesis and complement consumption has been correlated with disease activity. The presence of RF itself is associated with a more severe form of RA and the presence of extra-articular features.

Evidence exists (Janeway et al., J. Immunol., 138:1051 (1998); Rivera et al., Int. Immunol., 13: 1583-1593 (2001)) showing that B cells are highly efficient antigen-presenting cells (APC). RF-positive B cells may be particularly potent APCs, since their surface immunoglobulin would readily allow capture of any immune complexes regardless of the antigens present within them. Many antigens may thus be processed for presentation to T cells. In addition, it has been recently suggested that this may also allow RF-positive B cells to self-perpetuate. Edwards et al., Immunology, 97: 188-196 (1999).

For activation of T cells, two signals need to be delivered to the cell; one via the T-cell receptor (TCR), which recognizes the processed peptide in the presence of major histocompatibility complex (MHC) antigen, and a second, via co-stimulatory molecules. When activated, B cells express co-stimulatory molecules on their surface and can thus provide the second signal for T-cell activation and the generation of effector cells.

B cells may promote their own function as well as that of other cells by producing cytokines. Harris et al., Nat. Immunol., 1: 475-482 (2000). TNF-α, IL-1, lymphotoxin-α, and IL-10 are amongst some of the cytokines that B cells may produce in the RA synovium.

Although T-cell activation is considered to be a key component in the pathogenesis of RA, recent work using human synovium explants in severe combined immunodeficiency disorders (SCID) mice has demonstrated that T-cell activation and retention within the joint is critically dependent on the presence of B cells. Takemura et al., J. Immunol., 167: 4710-4718 (2001). The precise role of B cells in this is unclear, since other APCs did not appear to have the same effect on T cells.

Structural damage to joints is an important consequence of chronic synovial inflammation. Between 60% and 95% of patients with RA develop at least one radiographic erosion within 3-8 years of disease onset. Paulus et al., J. Rheumatol., 23: 801-805 (1996); Hulsmans et al., Arthritis Rheum., 43: 1927-1940 (2000). In early RA, the correlation between radiographic damage scores and functional capacity is weak, but after 8 years of disease, correlation coefficients can reach as high as 0.68. Scott et al., Rheumatology, 39:122-132 (2000). In 1,007 patients younger than age 60 years who had RA for at least four years, Wolfe et al. (Arthritis Rheum, 43 Suppl. 9:S403 (2000)) found a significant association among the rate of progression of the Larsen radiographic damage score (Larsen et al., Acta Radiol. Diagn. 18:481-491 (1977)), increasing Social Security disability status, and decreasing family income.

Diagnosis of RA may be according to current American College of Rheumatology (ACR) criteria and may include morning stiffness in and around the joints lasting for at least 1 hour before maximal improvement; arthritis of three or more joint areas: at least three joint areas have simultaneously had soft tissue swelling or fluid (not bony overgrowth alone) observed by a physician; the 14 possible joint areas (right and left) are proximal interphalangeal (PIP), metacarpophalangeal (MCP), wrist, elbow, knee, ankle, and metatarsophalangeal (MTP) joints; arthritis of hand joints: at least one joint area swollen as above in wrist, MCP, or PIP joint; symmetric arthritis: simultaneous involvement of the same joint areas (as in arthritis of three or more joint areas, above) on both sides of the body (bilateral involvement of PIP, MCP, or MTP joints is acceptable without absolute symmetry); rheumatoid nodules: subcutaneous nodules over bony prominences or extensor surfaces or in juxta-articular regions that are observed by a physician; serum rheumatoid factor: demonstration of abnormal amounts of serum rheumatoid factor by any method that has been positive in fewer than five percent of normal control patients; radiographic changes: radiographic changes typical of rheumatoid arthritis on posteroanterior hand and wrist X-rays, which must include erosions or unequivocal bony decalcification localized to or most marked adjacent to the involved joints (osteoarthritis changes alone do not qualify). Diagnosis of RA is typically made if a patient satisfies at least four of the above criteria.

Prevention or retardation of radiographic damage is one of the goals of RA treatment. Edmonds et al., Arthritis Rheum., 36:336-340 (1993). Controlled clinical trials of 6 or 12 months' duration have documented that the progression of radiographic damage scores was more rapid in the placebo group than in groups that received methotrexate (MTX) (Sharp et al., Arthritis Rheum., 43: 495-505 (2000)), leflunomide (Sharp et al., supra), sulfasalazine (SSZ) (Sharp et al., supra), prednisolone (Kirwan et al., N. Engl. J. Med., 333:142-146 (1995); Wassenberg et al., Arthritis Rheum, 42: Suppl 9:S243 (1999)), interleukin-1 receptor antagonist (Bresnihan et al., Arthritis Rheum, 41: 2196-2204 (1998)), or an infliximab/MTX combination. Lipsky et al., N. Eng. J. Med., 343: 1594-1604 (2000). Clinical trials have also documented that radiographic progression following treatment with etanercept was less rapid than that following treatment with MTX. Bathon et al., N. Engl. J. Med., 343:1586-1593 (2000). Other studies have evaluated radiographic progression in patients treated with corticosteroids (Joint Committee of the Medical Research Council and Nuffield Foundation, Ann Rheum. Dis., 19:331-337 (1960); Van Everdingen et al., Ann. Intern. Med., 136:1-12 (2002)), cyclosporin A (Priolo et al., J. Rheumatol., 24:2113-2118 (1997); Forre, Arthritis Rheum., 37:1506-1512 (1994)), MTX versus azathioprine (Jeurissen et al., Ann. Intern. Med., 114:999-1004 (1991)), MTX versus auranofin (Weinblatt et al., Arthritis Rheum., 36:613-619 (1993)), MTX (meta-analysis) (Alarcon et al., J. Rheumatol., 19:1868-1873 (1992)), hydroxychloroquine (HCQ) versus SSZ (Van der Heijde et al., Lancet, 1:1036-1038 (1989)), SSZ (Hannonen et al., Arthritis Rheum., 36:1501-1509 (1993)), the COBRA (Combinatietherapei Bij Reumatoide Artritis) combination of prednisolone, MTX, and SSZ (Boers et al., Lancet, 350:309-318 (1997); Landewe et al., Arthritis Rheum., 46: 347-356 (2002)), combinations of MTX, SSZ, and HCQ (O'Dell et al., N. Engl. J. Med., 334:1287-1291 (1996); Mottonen et al., Lancet, 353:1568-1573 (1999)), the combination of cyclophosphamide, azathioprine, and HCQ (Csuka et al., JAMA, 255:2315-2319 (1986)), and the combination of adalimumab with MTX. Keystone et al., Arthritis Rheum., 46 Suppl. 9:S205 (2002).

The FDA has now approved labeling claims that certain medications, e.g., leflunomide, etanercept, and infliximab, slow the progression of radiographic joint damage. These claims are based on the statistically significant differences in progression rates observed between randomly assigned treatment groups and control groups. However, the progression rates in individuals within the treatment and control groups overlap to a considerable extent. Therefore, despite significant differences between treatment groups, these data cannot be used to estimate the probability that a patient who is starting a treatment will have a favorable outcome with respect to progression of radiographic damage. Various methods have been suggested to categorize paired radiographs from individual patients as not progressive, e.g., damage scores of 0 at both time points, no increase in damage scores, no new joints with erosions, and a change in score not exceeding the smallest detectable difference (i.e., 95% confidence interval for the difference between repeated readings of the same radiograph). Lassere et al., J. Rheumatol., 26: 731-739 (1999).

Determining whether there has been increased structural damage in an individual patient during the interval between paired radiographs obtained at the beginning and end of a 6- or 12-month clinical trial has been difficult, for several reasons. The rate of radiographic damage is not uniform within a population of RA patients; a few patients may have rapidly progressing damage, but many may have little or no progression, especially if the tie interval is relatively short. The methods for scoring radiographic damage, e.g., Sharp (Sharp et al., Arthritis Rheum., 14: 706-720 (1971); Sharp et al., Arthritis Rheum., 28: 1326-1335 (1985)), Larsen (Larsen et al., Acta Radiol. Diagn., 18: 481-491 (1977)), and modifications of these methods (Van der Heijde, J. Rheumatol., 27: 261-263 (2000)), depend on the judgment and the interpretation of the reader as to what is real. Factors to determine are whether an apparent interruption of the subchondral cortical plate is real, or whether a decrease in the distance between the cortices on opposite sides of a joint is real, or is due to a slight change in the position of the joint relative to the film and the radiographic beam, to a change in radiographic exposure, or to some other technical factor.

Therefore, the recorded score is an approximation of the true damage, and for many subjects, the smallest detectable difference between repeat scores of the same radiographs is larger than the actual change that has occurred during the interval between the baseline and final radiographs. If the reader is blinded to the temporal sequence of the films, these unavoidable scoring errors may be in either direction, leading to apparent “healing” when the score decreases or to apparent rapid progression when reading error increases the difference between films. When the study involves a sufficiently large population of patients who have been randomly assigned to receive an effective treatment as compared with placebo, the positive and negative reading errors offset each other, and small but real differences between treatment groups can be detected.

The imprecision of the clinical measures that are used to quantitate RA disease activity has caused a similar problem. Statistically significant differences between certain outcome measures from clinical trials were not useful for estimating the probability of improvement for an individual who was starting the treatment. Paulus et al., Arthritis Rheum., 33:477-484 (1990). Attribution of individual improvement became practical with the creation of the American College of Rheumatology (ACR) 20% composite criteria for improvement (ACR20), which designated a patient as improved if there was 20% improvement in the tender and swollen joint counts and 20% improvement in at least three of five additional measures (pain, physical function, patient global health assessment, physician global health assessment, and acute-phase reactant levels). Felson et al., Arthritis Rheum., 38:727-735 (1995). All of these measures have large values for the smallest detectable difference, but by requiring simultaneous improvement in five of the seven aspects of the same process (disease activity), the randomness of the seven measurement errors is constrained, and it is easier to attribute real improvement to the individual.

In RA, joint damage is a prominent feature. Radiologic parameters of joint destruction are seen as a key outcome measure in descriptions of disease outcome. In the recent OMERACT (Outcome Measures in Rheumatology Clinical Trials) consensus meeting, radiology was chosen as part of the core set of outcome measures for longitudinal observational studies. Wolfe et al., Arthritis Rheum., 41 Supp 9: 5204 (1998) abstract. Radiology is also part of the WHO/ILAR (World Health Organization/International League of Associations for Rheumatology) required core set of measures for long-term clinical trials. Tugwell and Boers, J. Rheumatol., 20:528-530 (1993).

Available data on the outcome of radiologic damage in RA have been obtained in both short-term and long-term studies. In short-term studies of RA patients with recent-onset disease, radiographs obtained every six months showed that after an initial rapid progression, there was diminution of the progression rate of radiologic damage in the hands and feet after two to three years. Van der Heijde et al., Arthritis Rheum., 35: 26-34 (1992); Fex et al., Br. J. Rheumatol., 35: 1106-1115 (1996). In long-term studies with radiographs taken less frequently, a constant rate of progression was found, with relentless deterioration of damage up to 25 years of disease duration. Wolfe and Sharp, Arthritis Rheum., 41:1571-1582 (1998); Graudal et al., Arthritis Rheum., 41:1470-1480 (1998); Plant et al., J. Rheumatol., 25:417-426 (1998); Kaarela and Kautiainen, J. Rheumatol., 24:1285-1287 (1997). Whether these differences in radiographic progression pattern are due to differences in the scoring techniques is not clear.

The scoring systems used differ in the number of joints being scored, the presence of independent scores for erosions (ERO) and joint space narrowing (JSN), the maximum score per joint, and the weighing of a radiologic abnormality. As yet, there is no consensus on the scoring method of preference. During the first three years of follow-up in a cohort study of patients with early arthritis, JSN and ERO were found to differ in their contribution to the measured progression in radiologic damage of the hands and feet. Van der Heijde et al., Arthritis Rheum., 35:26-34 (1992). Furthermore, methods that independently score ERO and JSN, such as the Sharp and Kellgren scores, were found to be more sensitive to change in early RA than methods using an overall measure, such as the Larsen score. Plant et al., J. Rheumatol., 21:1808-1813 (1994); Cuchacovich et al., Arthritis Rheum., 35:736-739 (1992). The Sharp score is a very labor-intensive method. Van der Heijde, Baillieres Clin. Rheumatol., 10:435-453 (1996). In late or destructive RA, the Sharp and the Larsen methods were found to provide similar information. However, the sensitivity to change of the various scoring methods late in the disease has not yet been investigated, and it can be argued that the scoring methods that independently measure ERO and JSN provide useful information. Pincus et al., J. Rheumatol., 24:2106-2112 (1997). See also Drossaers-Bakker et al., Arthritis Rheum., 43:1465-1472 (2000), which compared the three radiologic scoring systems for the long-term assessment of RA.

Paulus et al., Arthritis Rheum., 50: 1083-1096 (2004) categorized radiographic joint damage as progressive or non-progressive in individuals with RA participating in clinical trials, and concluded that RA joint damage in an observational cohort can be classified as progressive or non-progressive with the use of a composite definition that includes a number of imprecise and related, but distinct, measures of structural joint damage. It appears that in day-to-day clinical management of an RA patient, an interval change between a pair of radiographs of at least five Sharp radiographic damage score units should be present before one considers the structural change to be real and uses it as the basis for a treatment decision.

Certain RA Therapeutic Agents

Initial therapy of RA typically involves administration of one or more of the following drugs: nonsteroidal antiinflammatory drugs (NSAIDs), e.g., acetylsalicylic acid (e.g., aspirin), ibuprofen (Motrin), naproxen (Naprosyn), indomethacin (Indocin), nabumetone (Relafen), tolmetin (Tolectin); glucocorticoid (via joint injection); and low-dose prednisone. See “Guidelines for the management of rheumatoid arthritis,” Arthritis & Rheumatism 46(2): 328-346 (February, 2002). The majority of patients with newly diagnosed RA are started with disease-modifying antirheumatic drug (DMARD) therapy within 3 months of diagnosis. DMARDs commonly used in RA are hydroxychloroquine, sulfasalazine, methotrexate (plus oral and subcutaneous methotrexate), leflunomide, azathioprine, D-penicillamine, Gold (oral), Gold (intramuscular), minocycline, cyclosporine, Staphylococcal protein A immunoadsorption. In certain instances, patients are treated with immunomodulating agents such as azathioprine or cyclophosphamide. Additional RA therapeutic agents include an anti-cytokine agent (e.g., anti-tumor necrosis factor α, anti-interleukin-1-receptor (e.g., anakinra), anti-interleukin 10, anti-interleukin 6 receptor, anti-interleukin 6, anti-interferon alpha, anti-B-lymphocyte stimulator), an inhibitor of costimulation (e.g., anti-CD154, CTLA4-Ig (e.g., abatacept)).

In certain instances, TNFα inhibitors have been used for therapy of RA. Exemplary TNFα inhibitors include etanercept (sold under the trade name ENBREL®), infliximab (sold under the trade name REMICADE®), adalimumab (sold under the trade name HUMIRA®), golimumab (sold under the trade name SIMPONI™) and certolizumab pegol (sold under the trade name CIMZIA®).

Etanercept (sold under the trade name ENBREL®) is an injectable drug approved in the U.S. for therapy of active RA. Etanercept binds to TNFα and serves to remove most TNFα from joints and blood, thereby preventing TNFα from promoting inflammation and other symptoms of rheumatoid arthritis. Etanercept is an “immunoadhesin” fusion protein consisting of the extracellular ligand binding portion of the human 75 kD (p75) tumor necrosis factor receptor (TNFR) linked to the Fc portion of a human IgG1. The drug has been associated with negative side effects including serious infections and sepsis, and nervous system disorders such as multiple sclerosis (MS). See, e.g., www.remicade-infliximab.com/pages/enbrel_embrel.html.

Infliximab, sold under the trade name REMICADE®, is an immune-suppressing drug prescribed to treat RA and Crohn's disease. Infliximab is a chimeric monoclonal antibody that binds to TNFα and reduces inflammation in the body by targeting and binding to TNFα which produces inflammation. Infliximab has been linked to certain fatal reactions such as heart failure and infections including tuberculosis as well as demyelination resulting in MS. See, e.g., www.remicade-infliximab.com.

In 2002, Abbott Laboratories received FDA approval to market adalimumab (sold under the trade name HUMIRA®), previously known as D2E7. Adalimumab is a human monoclonal antibody that binds to TNFα and is approved for reducing the signs and symptoms and inhibiting the progression of structural damage in adults with moderately to severely active RA who have had insufficient response to one or more traditional disease modifying DMARDs.

In April 2009, Centocor Ortho Biotech Inc. received FDA approval to market golimumab (sold under the trade name SIMPONI™) for patients with moderate to severe RA, psoriatic arthritis, and ankylosing spondylitis. Golimumab is a human IgG1κ monoclonal antibody specific for human TNFα and which is self-administered by patients subcutaneously once every month. Golimumab binds to both soluble and transmembrane bioactive forms of TNFα. Similar to other agents that inhibit TNFα, golimumab has been associated with certain adverse events such as risk of infection, including serious and life-threatening fungal infections.

In May 2009, certolizumab pegol (sold under the trade name CIMZIA®) was approved by the FDA for treatment of patients with RA. It is administered by a healthcare professional by subcutaneous injection every two weeks during induction and then every four weeks during maintenance. Certolizumab pegol is a recombinant, humanized antibody Fab′ fragment, with specificity for human TNFα, conjugated to an approximately 40 kDa polyethylene glycol (PEG2MAL40K). Certolizumab pegol has also been associated with certain safety risks such as increased risk of serious infection, similar to other TNFα inhibitors.

In certain instances, the rituximab antibody (sold under the trade name RITUXAN®) has been used as a therapy for RA. Rituximab is a genetically engineered chimeric murine/human monoclonal antibody directed against the CD20 antigen. Rituximab is the antibody called “C2B8” in U.S. Pat. No. 5,736,137 issued Apr. 7, 1998 (Anderson et al.).

Another anti-CD20 antibody is ocrelizumab. Ocrelizumab is a humanized variant of an anti-CD20 antibody, 2H7. Such humanized 2H7 variants are described, for example, in International Publication No. WO 2004/056312 (International Application No. PCT/US2003/040426).

RA therapeutic agents having B-cell antagonist activity can be identified, for example, by screening compounds for certain biological properties. For example, a method of screening can be employed as described in Sundberg et al., Cancer Research 66, 1775-1782 (2006) wherein a compound was screened for inhibition of B-cell proliferation by targeting c-myc protein for rapid and specific degradation. See also Mackay et al., Annual Review of Immunology, 21: 231-264 (2003) regarding BAFF, APRIL, and a tutorial on B-cell survival and screening, and Thangarajh et al., Scandinavian J. Immunol., 65(1):92 (2007) on B-cell proliferation and APRIL. In addition, Sakurai et al., European J. Immunol., 37(1):110 (2007) discloses that TACT attenuates antibody production co-stimulated by BAFF-R and CD40. Further, Acosta-Rodriguez et al., European J. Immunol., 37(4):990 (2007) discloses that BAFF and LPS cooperate to induce B cells to become susceptible to CD95/Fas-mediated cell death. Further screening methods can be found in Martin and Chan, “B Cell Immunobiology in Disease: Evolving Concepts from the Clinic Annual Review of Immunology,” 24:467-496 (2006), Pillai et al., “Marginal Zone B Cells” Annual Review of Immunology, 23:161-196 (2005), and Hardy and Hayakawa, “B Cell Development Pathways,” Annual Review of Immunology, 19:595-621 (2001). From these and other references the skilled artisan can screen for the appropriate antagonists. Microarrays can be used for this purpose (Hagmann, Science, 290:82-83 (2000)), as well as RNA interference (RNAi) (Ngo et al., Nature, 441:106-110 (2006)).

B-cell antagonists included within the scope of the present invention include antibodies, synthetic or native-sequence peptides, immunoadhesins, and small-molecule antagonists that bind to a B-cell surface marker or a B-cell specific survival or proliferation factor, optionally conjugated with or fused to another molecule. In certain embodiments, the antagonist comprises an antibody or immunoadhesin. It includes BLyS antagonists such as immunoadhesins, including, but not limited to, anti-CD23 (e.g., lumiliximab), anti-CD20, anti-CD22, or anti-BR3 antibodies, APRIL antagonists, and/or BLyS immunoadhesins. In certain embodiments, the BLyS immunoadhesin is selected from BR3 immunoadhesin comprising the extracellular domain of BR3, TACI immunoadhesin comprising the extracellular domain of TACI, and BCMA immunoadhesin comprising the extracellular domain of BCMA. Certain embodiments of BR3 immunoadhesin include hBR3-Fc as described in WO 2005/00351, U.S. Pat. Pub. No. 2005/0095243, U.S. Pat. Pub. No. 2005/0163775 and WO 2006/068867. In certain embodiments, the BLyS antagonist is an anti-BLyS antibody, wherein the anti-BLyS antibody binds BLyS within a region of BLyS comprising residues 162-275, or an anti-BR3 antibody, wherein the anti-BR3 antibody binds BR3 in a region comprising residues 23-38 of human BR3. In certain embodiments, the immunoadhesins are selected from TACI-Ig (atacicept) and BR3-Ig. In certain embodiments, the B-cell antagonist is to CD20, CD22, BAFF, or APRIL. In certain such embodiments, the antagonist is an antibody or TACI-Ig.

The CD22 antigen, or CD22, also known as BL-CAM or Lyb8, is a type 1 integral membrane glycoprotein with molecular weight of about 130 (reduced) to 140 kD (unreduced). It is expressed in both the cytoplasm and cell membrane of B-lymphocytes. CD22 antigen appears early in B-cell lymphocyte differentiation at approximately the same stage as the CD19 antigen. Unlike certain other B-cell markers, CD22 membrane expression is limited to the late differentiation stages comprised between mature B cells (CD22+) and plasma cells (CD22−). The CD22 antigen is described, for example, in Wilson et al., J. Exp. Med., 173:137 (1991) and Wilson et al., J. Immunol., 150:5013 (1993).

Certain exemplary anti-CD22 antibodies include those described in EP 1,476,120 (Tedder and Tuscano), EP 1,485,130 (Tedder), and EP 1,504,035 (Popplewell et al.), as well as those described in U.S. Pat. Pub. No. 2004/0258682 (Leung et al.), U.S. Pat. No. 5,484,892 (Dana-Farber), U.S. Pat. No. 6,183,744 (Immunomedics, epratuzumab), and U.S. Pat. No. 7,074,403 (Goldenberg and Hansen).

BLyS (also known as BAFF, TALL-1, THANK, TNFSF13B, or zTNF4) is a member of the TNF1 ligand superfamily that is essential for B-cell survival and maturation. BAFF overexpression in transgenic mice leads to B-cell hyperplasia and development of severe autoimmune disease (Mackay et al., J. Exp. Med., 190:1697-1710 (1999); Gross et al., Nature, 404:995-999 (2000); Khare et al., Proc. Natl. Acad. Sci. U.S.A, 97:3370-3375 (2000)). BAFF levels are elevated in human patients with a variety of autoimmune disorders, such as SLE, RA, and Sjögren's syndrome (Cheema et al., Arthritis Rheum., 44:1313-1319 (2001); Groom et al, J. Clin. Invest., 109:59-68 (2002); Zhang et al., J. Immunol., 166:6-10 (2001)). Furthermore, BAFF levels correlate with disease severity, suggesting that BAFF can play a direct role in the pathogenesis of these illnesses. BAFF acts on B cells by binding to three members of the TNF receptor superfamily, TACI, BCMA, and BR3 (also known as BAFF-R) (Gross et al., supra; Thompson et al., Science, 293:2108-2111 (2001); Yan et al., Curr. Biol. 11:1547-1552 (2001); Yan et al., Nat. Immunol., 1:37-41 (2000); Schiemann et al., Science, 293:2111-2114 (2001)).

Of the three, only BR3 is specific for BAFF; the other two also bind the related TNF family member, A proliferation-inducing ligand (APRIL). Comparison of the phenotypes of BAFF and receptor knockout or mutant mice indicates that signaling through BR3 mediates the B-cell survival functions of BAFF (Thompson et al., supra; Yan et al., supra, 2001; Schiemann et al., supra). In contrast, TACI ap-pears to act as an inhibitory receptor (Yan, Nat. Immunol., 2:638-643 (2001)), while the role of BCMA is unclear (Schiemann et al., supra). US 2007/0071760 discloses treating B-cell malignancies using a TACI-Ig fusion molecule in an amount sufficient to suppress proliferation-inducing functions of BlyS and APRIL.

BR3 is a 184-residue type III transmembrane protein expressed on the surface of B cells (Thompson et al., supra; Yan, Nat. Immun., supra). The intracellular region bears no sequence similarity to known structural domains or protein-protein interaction motifs. Nevertheless, BAFF-induced signaling through BR3 results in processing of the transcription factor NF-B2/p100 to p52 (Claudio et al., Nat. Immunol., 3:958-965 (2002); Kayagaki et al., Immunity, 10:515-524 (2002)). The extracellular domain (ECD) of BR3 is also divergent. TNFR family members are usually characterized by the presence of multiple cysteine-rich domains (CRDs) in their extracellular region; each CRD is typically composed of about 40 residues stabilized by six cysteines in three disulfide bonds. Conventional members of this family make contacts with ligand through two CRDs interacting with two distinct patches on the ligand surface (Bodmer et al., Trends Biochem. Sci., 27:19-26 (2002)). However, the BR3 ECD contains only four cysteine residues, capable of forming a partial CRD at most, raising the question of how such a small receptor imparts high-affinity ligand binding.

It has been shown that the BAFF-binding domain of BR3 resides within a 26-residue core region (Kayagaki et al., supra). Six BR3 residues, when structured within a β-hairpin peptide (bhpBR3), were sufficient to confer BAFF binding and block BR3-mediated signaling. Others have reported polypeptides purported to interact with BAFF (e.g., WO 2002/24909, WO 2003/035846, WO 2002/16312, and WO 2002/02641).

Loss of function and radiographic change occur early in the course of the disease. These changes can be delayed or prevented with the use of certain DMARDs. Although several DMARDs are initially clinically effective and well tolerated, many of these drugs become less effective or exhibit increased toxicity over time. Based on its efficacy and tolerability, MTX has become the standard therapy by which other treatments are measured. Bathon et al., N. Eng. J. Med., 343:1586-1593 (2000); Albert et al., J. Rheumatol., 27:644-652 (2000).

Recent studies have examined radiographic progression in patients with late-stage RA who have taken leflunomide, MTX, or placebo (Strand et al., Arch. Intern. Med., 159:2542-2550 (1999)) as well as patients who have taken infliximab plus MTX or placebo plus MTX following a partial response to MTX. Lipsky et al., N. Engl. J. Med., 343:1594-1602 (2000); Maini et al., Lancet, 354:1932-1939 (1999). In the first year of the ENBREL™ ERA (early RA) trial, etanercept was shown to be significantly more effective than MTX in improving signs and symptoms of disease and in inhibiting radiographic progression. Bathon et al., N. Eng. J. Med., 343:1586-1593 (2000). Genovese et al., Arthritis Rheum. 46:1443-1450 (2002) reports results from the second year of the study, concluding that etanercept as monotherapy was safe and superior to MTX in reducing disease activity, arresting structural damage, and decreasing disability over two years in patients with early aggressive RA. Also studied was the safety and clinical activity of ocrelizumab (a humanized antibody targeting C D20+B cells) in combination with MTX in moderate-to-severe RA patients (Ph I/II ACTION study). Genovese et al., Arthritis Rheum., 54(9):S66-S67 (September 2006).

Further, reduction in radiographic progression in the hands and feet was observed in patients with early RA after receiving infliximab in combination with MTX. Van der Heijde et al., Annals Rheumatic Diseases, 64:417 (2005). Patients with early RA achieved a clinically meaningful and sustained improvement in physical function after treatment with infliximab. Smolen et al., Annals Rheumatic Diseases, 64:418-419 (2005).

The effect of infliximab therapy on bone mineral density in patients with ankylosing spondylitis (AS) resulting from a randomized, placebo-controlled trial named ASSERT) is reported by Van der Heijde et al., Annals Rheumatic Diseases, 64:319 (2005). The ASSERT trial showed that infliximab improved fatigue and pain in patients with AS. Van der Heijde et al., Annals Rheumatic Diseases, 64:318-319 (2005). The efficacy and safety of infliximab in AS patients treated according to ASSERT are described by van der Heijde et al., Arthritis Rheum., 52:582-591 (2005). The authors conclude that infliximab was well tolerated and effective in a large cohort of patients with AS during a 24-week study period. In addition, the effect of infliximab therapy on spinal inflammation was assessed by magnetic resonance imaging in a randomized, placebo-controlled trial of 279 patients with AS. Van der Heijde et al., Annals Rheumatic Diseases, 64:317 (2005). The manner in which the treatment effect on spinal radiographic progression in patients with AS should be measured is addressed by van der Heijde et al., Arthritis Rheum. 52:1979-1985 (2005).

The results of radiographic analyses of the infliximab multinational PsA controlled trial (IMPACT) after one year are reported by Antoni et al., Annals Rheumatic Diseases 64:107 (2005). Evidence of radiographic benefit of treatment with infliximab plus MTX in RA patients who had no clinical improvement, with a detailed subanalysis of data from the anti-TNF trial in RA with concomitant therapy study, is reported by Smolen et al., Arthritis Rheum. 52:1020-1030 (2005). Radiographic progression (as measured by mean change in modified Sharp/van der Heijde score) was much greater in patients receiving MTX plus placebo than in patients receiving infliximab plus MTX. The authors conclude that even in patients without clinical improvement, treatment with infliximab plus MTX provided significant benefit with regard to the destructive process, suggesting that in such patients these two measures of disease are dissociated. The association between baseline radiographic damage and improvement in physical function after treatment of patients having RA with infliximab is described by Breedveld et al., Annals Rheumatic Diseases, 64:52-55 (2005). Structural damage was assessed using the van der Heijde modification of the Sharp score. The authors conclude that greater joint damage at baseline was associated with poorer physical function at baseline and less improvement in physical function after treatment, underlining the importance of early intervention to slow the progression of joint destruction.

Rheumatoid Arthritis Molecular Biomarkers

A number of investigators have carried out microarray gene expression profiling studies of synovial tissue isolated from RA patients. The published studies include van der Pouw Kraan T C et al., Discovery of distinctive gene expression profiles in rheumatoid synovium using cDNA microarray technology: evidence for the existence of multiple pathways of tissue destruction and repair, Genes Immun April; 4(3):187-96 (2003); van der Pouw Kraan T C, et al., Rheumatoid arthritis is a heterogeneous disease: evidence for differences in the activation of the STAT-1 pathway between rheumatoid tissues, Arthritis Rheum August;48(8):2132-45 (2003); Finis K et al., Analysis of pigmented villonodular synovitis with genome-wide complementary DNA microarray and tissue array technology reveals insight into potential novel therapeutic approaches, Arthritis Rheum March; 54(3):1009-19 (2006); Lindberg J, et al., Effect of infliximab on mRNA expression profiles in synovial tissue of rheumatoid arthritis patients, Arthritis Res Ther. 8(6):R179 (2006); van der Pouw Kraan T C et al., Responsiveness to anti-tumour necrosis factor alpha therapy is related to pre-treatment tissue inflammation levels in rheumatoid arthritis patients, Ann Rheum Dis. April; 67(4):563-6 (2008); Huber R et al., Identification of intra-group, inter-individual, and gene-specific variances in mRNA expression profiles in the rheumatoid arthritis synovial membrane, Arthritis Res Ther 10(4):R98 (2008); Badot V et al., Gene expression profiling in the synovium identifies a predictive signature of absence of response to adalimumab therapy in rheumatoid arthritis, Arthritis Res Ther. 11(2):R57 (2009), Epub 2009 Apr. 23.

General Techniques

The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology, which are within the skill of the art. Such techniques are explained fully in the literature, such as, “Molecular Cloning: A Laboratory Manual”, second edition (Sambrook et al., 1989); “Oligonucleotide Synthesis” (M. J. Gait, ed., 1984); “Animal Cell Culture” (R. I. Freshney, ed., 1987); “Methods in Enzymology” (Academic Press, Inc.); “Current Protocols in Molecular Biology” (F. M. Ausubel et al., eds., 1987, and periodic updates); “PCR: The Polymerase Chain Reaction”, (Mullis et al., eds., 1994).

Primers, oligonucleotides and polynucleotides employed in the present invention can be generated using standard techniques known in the art.

Gene expression signatures associated with RA and certain subtypes of RA are provided herein. These signatures constitute biomarkers for RA and/or subtypes of RA, and/or predispose or contribute to development, persistence and/or progression of RA. Accordingly, the invention disclosed herein is useful in a variety of settings, e.g., in methods and compositions related to RA diagnosis and therapy.

Detection of Gene Expression Levels

Nucleic acid, according to any of the methods described herein may be RNA transcribed from genomic DNA or cDNA generated from RNA. Nucleic acid may be derived from a vertebrate, e.g., a mammal. A nucleic acid is said to be “derived from” a particular source if it is obtained directly from that source or if it is a copy of a nucleic acid found in that source.

Nucleic acid includes copies of the nucleic acid, e.g., copies that result from amplification. Amplification may be desirable in certain instances, e.g., in order to obtain a desired amount of material for detecting variations. The amplicons may then be subjected to a variation detection method, such as those described below, to determine expression of certain genes.

A microarray is a multiplex technology that typically uses an arrayed series of thousands of nucleic acid probes to hybridize with, e.g, a cDNA or cRNA sample under high-stringency conditions. Probe-target hybridization is typically detected and quantified by detection of fluorophore-, silver-, or chemiluminescence-labeled targets to determine relative abundance of nucleic acid sequences in the target. In typical microarrays, the probes are attached to a solid surface by a covalent bond to a chemical matrix (via epoxy-silane, amino-silane, lysine, polyacrylamide or others). The solid surface is for example, glass, a silicon chip, or microscopic beads. Various microarrays are commercially available, including those manufactured, for example, by Affymetrix, Inc. and Illumina, Inc.

A biological sample may be obtained using certain methods known to those skilled in the art. Biological samples may be obtained from vertebrate animals, and in particular, mammals. In certain instances, a biological sample is synovial tissue, serum or peripheral blood mononuclear cells (PBMC). By screening such body samples, a simple early diagnosis can be achieved for diseases such as RA. In addition, the progress of therapy can be monitored more easily by testing such body samples for variations in expression levels of target nucleic acids (or encoded polypeptides).

Subsequent to the determination that a subject, or the tissue or cell sample comprises a gene expression signature disclosed herein, it is contemplated that an effective amount of an appropriate RA therapeutic agent may be administered to the subject to treat the RA in the subject. Clinical diagnosis in mammals of the various pathological conditions described herein can be made by the skilled practitioner. Clinical diagnostic techniques are available in the art which allow, e.g., for the diagnosis or detection of RA in a mammal.

A RA therapeutic agent can be administered in accordance with known methods, such as intravenous administration as a bolus or by continuous infusion over a period of time, by intramuscular, intraperitoneal, intracerobrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral, topical, or inhalation routes. Optionally, administration may be performed through mini-pump infusion using various commercially available devices.

Kits

For use in the applications described or suggested herein, kits or articles of manufacture are also provided. Such kits may comprise a carrier means being compartmentalized to receive in close confinement one or more container means such as vials, tubes, and the like, each of the container means comprising one of the separate elements to be used in the method. For example, one of the container means may comprise a probe that is or can be detectably labeled. Such probe may be a polynucleotide specific for a polynucleotide comprising one or more genes of a gene expression signature. Where the kit utilizes nucleic acid hybridization to detect the target nucleic acid, the kit may also have containers containing nucleotide(s) for amplification of the target nucleic acid sequence and/or a container comprising a reporter means, such as a biotin-binding protein, such as avidin or streptavidin, bound to a reporter molecule, such as an enzymatic, florescent, or radioisotope label.

Kits will typically comprise the container described above and one or more other containers comprising materials desirable from a commercial and user standpoint, including buffers, diluents, filters, needles, syringes, and package inserts with instructions for use. A label may be present on the container to indicate that the composition is used for a specific therapy or non-therapeutic application, and may also indicate directions for either in vivo or in vitro use, such as those described above. Other optional components in the kit include one or more buffers (e.g., block buffer, wash buffer, substrate buffer, etc), other reagents such as substrate (e.g., chromogen) which is chemically altered by an enzymatic label, epitope retrieval solution, control samples (positive and/or negative controls), control slide(s) etc.

Methods of Marketing

The invention herein also encompasses a method for marketing a RA therapeutic agent or a pharmaceutically acceptable composition thereof comprising promoting to, instructing, and/or specifying to a target audience, the use of the agent or pharmaceutical composition thereof for treating a patient or patient population with RA from which a sample has been obtained showing the presence of a genetic variation as disclosed herein.

Marketing is generally paid communication through a non-personal medium in which the sponsor is identified and the message is controlled. Marketing for purposes herein includes publicity, public relations, product placement, sponsorship, underwriting, and sales promotion. This term also includes sponsored informational public notices appearing in any of the print communications media designed to appeal to a mass audience to persuade, inform, promote, motivate, or otherwise modify behavior toward a favorable pattern of purchasing, supporting, or approving the invention herein.

The marketing of the diagnostic method herein may be accomplished by any means. Examples of marketing media used to deliver these messages include television, radio, movies, magazines, newspapers, the internet, and billboards, including commercials, which are messages appearing in the broadcast media.

The type of marketing used will depend on many factors, for example, on the nature of the target audience to be reached, e.g., hospitals, insurance companies, clinics, doctors, nurses, and patients, as well as cost considerations and the relevant jurisdictional laws and regulations governing marketing of medicaments and diagnostics. The marketing may be individualized or customized based on user characterizations defined by service interaction and/or other data such as user demographics and geographical location.

EXAMPLES

The following are examples of the methods and compositions of the invention. It is understood that various other embodiments may be practiced, given the general description provided above.

Example 1 Methods and Subjects Subjects and Synovial Biopsies

All procedures involving specimens obtained from human subjects were performed under a protocol approved by the University of Michigan Institutional Review Board. Human synovial tissues were obtained by synovectomy from affected joints in patients diagnosed with RA based upon the presence of at least four of the seven criteria developed by the American College of Rheumatology for RA (Arnett, F. C., et al., Arthritis Rheum., 31: 315-324 (1988)). Excised tissues were immediately snap-frozen in liquid nitrogen and stored at −80° C. For matched histology sections, samples were brought briefly to −20° C., cryostat sectioned and immediately brought back to −80° C. Frozen samples were homogenized in Qiagen brand RLT and RNA was isolated according to the manufacturers recommended protocol (Qiagen, Valencia, Calif.)

Methods

Microarray Hybridization

The methods for preparation of cRNA and for array hybridization were provided by Affymetrix, Inc. (Santa Clara, Calif.). Briefly, 3 μg of total RNA was converted into double-stranded cDNA using a cDNA synthesis kit, SuperScript Choice (Invitrogen, Carlsbad, Calif.) and a T7-(dT)₂₄ oligomer primer (Biosearch Technologies, Inc., Novato, Calif.). Double-stranded cDNA was purified using affinity resin Sample Cleanup Module Kit (Affymetrix, Inc.) and then ethanol precipitated. Labeled cRNA was generated from the cDNA by using a T7 RNA polymerase and biotin-labeled nucleotide in an in vitro transcription reagents (Enzo Diagnostics, Inc., Farmingdale, N.Y.). The labeled cRNA was purified using Affymetrix Sample Cleanup Module Kit. The amount of labeled cRNA was determined by measuring absorbance at 260 nm and using the convention that 1 OD at 260 nm corresponds to 40 μg/ml of RNA. Fifteen micrograms of labeled cRNA was fragmented by incubating at 94° C. for 30 min in 40 mM Tris-acetate pH 8.1, 100 mM potassium acetate and 30 mM magnesium acetate. Samples were then hybridized to GeneChip® Human Genome U133 Plus 2.0 Arrays (Affymetrix, Inc.) at 45° C. for 19 hours in a rotisserie oven set at 60 rpm. Arrays were washed and stained in the Affymetrix Fluidics station and scanned on GeneChip® scanner 3000. Data analysis was performed using the Affymetrix GeneChip® operating system and analysis software.

Histopathology and Immunohistochemistry

Stains were performed on 5-μm-thick frozen sections of human synovial tissue fixed in acetone. Some sections were stained with hematoxylin and eosin for histologic evaluation. Other sections were blocked in 10% serum for 30 minutes and stained for the detection of cells expressing the following lineage markers (CD20—mouse anti-human clone L26, 5 μg/ml, Dako; CD3—rabbit anti-human antibody SP7, 1:200 dilution, NeoMarkers; and CD68—mouse anti-human clone KP-1, 2.5 μg/ml Dako). All immunohistochemical stains were detected with species specific, biotinylated secondary antibodies and 3,3′-diaminobenzidine (DAB).

Statistical Analyses

Statistical analyses of microarray data was performed with the open-source tools available in the statistical programming environment, R (available at the URL: cran(dot)r-project(dot)org) and the commercially available Spotfire Decision Site (TIBCO Software Inc, Palo Alto, Calif.). Identification of molecular subytpes was performed by multi-scale bootstrap resampling using the open-source R package, Pvclust (Suzuki, R. and Shimodaira, H., Bioinformatics, 22(12), 1540-1542 (2006)). Heatmap visualizations and identification of differentially expressed genes was performed using analysis of variance provided by Spotfire. Identification of pathways significantly over-represented within each subtype was performed using CoPub, following the developers' recommended protocol (Frijters, R. et al., Nucleic Acids Res. 36:W406-W410 (Web server issue doi:10.1093/nar/gkn215) (2008)); available at the URL: services(dot)nbic(dot)nl(slash)cgi-bin(slash)copub(slash)microarray_analysis(dot)pl. Briefly, Affymetrix probeset identifiers that were specifically upregulated within each subtype (˜1000 top ranked probesets) were uploaded to the web-server. The GeneChip® Human Genome U133A Plus 2.0 Array (Affymetrix, Inc.) was selected as the background data set, the search category was limited to biological processes and all calculation settings were left at their defaults. The resulting data was saved to a personal computer and formatted for comparative heatmap visualization in Spotfire.

Identification of Classifiers: Molecular Phenotype Training and Testing

Using the filtered expression data set consisting of 20,776 probes and the class labels we sought to build a series of two-class and multiclass classification models which could distinguish (i) each putative patient subclass from the other three subtypes or (ii) mutually distinguish all four subclasses from each other, respectively. We refer to such classification models herein as “classifiers.” In the case where multiple samples were available from the same patient, one sample from that patient was selected at random to enter into the model. Variable (probe) selection and model training was performed using the CMA package (Slawski et al., BMC Bioinformatics 9:439 (2008)). In the case of the two-class models, variable selection was performed by ranking each probe's association with a given class label according to either the absolute value of its two-sample t-statistic or its robust Wilcoxon statistic. For the multiclass model, each probe was ranked by the values of its one-way F-statistic or its robust Kruskall-Wallis test statistic across all four putative classes. The values of the test statistics were recorded over N=48 rounds of leave-one-out cross-validation (LOOCV), or, when the class sizes were deemed large enough, i.e., for the F2, L and M two-class models, over 100 repeated rounds of 5-fold cross-validation. For each model and choice of test statistic, and at each round of cross-validation, a list of the top 20 probes with the largest, most significant values of their test statistic was retained. A probe-specific voting-based variable importance measure was created in which the number (or fraction) of rounds of cross-validation a probe appeared in the list of the top 20 most strongly associated probes was calculated.

Performing linear discriminant analysis (LDA) in the CMA package, an estimated class label, obtained from using these specific 48 patient samples, could be compared to the original estimated labels of the clustering results. As a sanity check, the variable selection and LDA steps were repeated using permuted class labels, resulting in increased rates of misclassification.

Publicly available independent test data on a two-color microarray platform (Lindberg et al., PLoS One 5(6):e11310 (2010)) were used to assess the robustness of the models constructed from the training data. For each RA patient two-two-way model and for the multiclass model, the set of unique probes, aggregated over choice of parametric or robust test statistic, which ever appeared in a given round of cross-validation's list of top 20 probes, was applied to a LDA model on the training data using the MASS package in R (Venables, W. N. & Ripley, B. D. (2002) Modern Applied Statistics with S. Fourth Edition. Springer, New York. ISBN 0-387-95457-0). Using LOOCV, new, predicted class labels were obtained by applying the LDA model built on the training data to the new test set data. Probes between the two data sets were linked by their unique Entrez Gene identification number. In the case where multiple probes in either data set mapped to a given Entrez Gene number, a unique probe was selected to represent a given gene. In the original Affymetrix training data, the probe with the highest variable importance score over rounds of LOOCV was selected. In the case of ties, one probe was chosen at random. Unique representative probes in the test data were also selected at random. Missing data in the test data set were imputed using the median expression value for that probe. Prior to performing LDA, both the training and test data were centered and scaled to place them on more equal footing. The classifiers for each of the four molecular phenotypes are provided below.

Identification of Molecular Phenotypes (Subtypes)

Gene expression microarray experiments on synovial tissues isolated from patients with RA were carried out, for example, to assess gene expression patterns as a basis to advance understanding of the molecular pathways important in RA pathogenesis and progression as well as to identify potential therapeutic targets and biomarkers for diagnostic and prognostic purposes. Gene expression microarray experiments on 81 synovial tissue samples, excised from 50 RA patients, were carried out using a whole genome expression array, the GeneChip® Human Genome U133 Plus 2.0 Array (Affymetrix, Inc.). Expression data was normalized using manufacturer provided software, MAS5, standardized to 500, log transformed and z-scored. A probe was included in the analysis if it had a minimum expression of at least 100 and it varied by 1.5 standard deviations in at least 5 samples relative to probe's mean expression level across all samples. This assessment yielded 20,776 probes, which were randomly sampled with replacement for 10,000 iterations and clustered using correlation as the distance metric and average linkage for agglomeration. The resulting dendrogam shown in FIG. 1 depicts the sample clustering and the resulting bootstrapped branch support values.

We analyzed the heatmap resulting from the microarray experiment (FIG. 2) and identified four molecular subtypes of RA based on relative expression levels of genes differentially expressed between the different RA molecular subtypes. We assigned samples to four distinct groups inferred from the bootstrapped dendrogram (FIG. 2). Analysis of variance on log transformed expression data identified differentially expressed genes within in each group. Hierarchical clustering was performed on statistically significant probes (5501 probes with p<1.0E-6). Expression data was z-score normalized for visualization.

As shown in FIG. 2, the largest grouping of samples (45%) defined the molecular subtype described herein as lymphoid-rich (L) (FIG. 2, the branches at the top of the dendrogram labeled “L” at the top of figure and corresponding box shown within the heatmap, 87% bootstrap support). These samples shared extensive lymphoid infiltration and follicle-like lymphoid clusters (each p<0.01). The gene expression signature of this group of samples revealed a pattern characteristic of B cells, plasma cells, T cells, and macrophages and implicated certain pathways including B and T cell activation, isotype switching, Ig secretion and cytokine production. Table 5 provides a list of probe sets (and associated genes) that are associated with L subtype. These were identified from the microarray data using the following criteria: (1) Fold change≧1.5 within the L subtype; (2) t-test p-value≧0.0001; and (3) annotated as belonging to at least one of the following molecular categories of proteins: secreted, plasma membrane, kinase, G-coupled protein receptor, phosphatase, nuclear receptor, ion channel, E3-ligase, de-ubiquitinating enzyme.

Table 1 below shows a subset of certain of these probe sets (and associated genes) from Table 5 that have been identified as therapeutic targets and biomarkers of the L subtype. The genes identified in Table 1 encode proteins that share the properties of surface expression and secretion. Proteins having those properties can, in certain instances, be targeted with, for example, monoclonal antibodies and in that case are considered therapeutic targets. Secreted proteins and products cleaved from the cell membrane can, in certain cases, be measured and in that case are considered biomarkers.

TABLE 1 Certain L subtype therapeutic targets and biomarkers. NETAFFX: NETAFFX: Unison: Fold ProbeID GeneTitle GeneSymbol Protcomp Change p-value 210356_x_at membrane-spanning MS4A1 Plasma membrane 5.182001696 1.07E−13 4-domains, subfamily A, member 1 217418_x_at membrane-spanning MS4A1 Plasma membrane 5.708081511 6.62E−13 4-domains, subfamily A, member 1 228592_at Membrane-spanning MS4A1 Plasma membrane 8.475244795 1.06E−12 4-domains, subfamily A, member 1 228599_at Membrane-spanning MS4A1 Plasma membrane 4.628458246 4.08E−13 4-domains, subfamily A, member 1 221331_x_at cytotoxic T- CTLA4 Plasma membrane 2.342826855 7.67E−11 lymphocyte- associated protein 4 231794_at cytotoxic T- CTLA4 Plasma membrane 1.785965453 7.40E−08 lymphocyte- associated protein 4 234362_s_at cytotoxic T- CTLA4 Plasma membrane 1.971205062 2.69E−12 lymphocyte- associated protein 4 236341_at cytotoxic T- CTLA4 Plasma membrane 2.398839174 6.75E−09 lymphocyte- associated protein 4 213539_at CD3D antigen, delta CD3D Plasma membrane 2.45243047 1.43E−08 polypeptide (TiT3 complex) 205456_at CD3E antigen, CD3E Plasma membrane 1.915380475 2.79E−10 epsilon polypeptide (TiT3 complex) 206804_at CD3G antigen, CD3G Plasma membrane 3.056708104 6.56E−06 gamma polypeptide (TiT3 complex) 210031_at CD3Z antigen, zeta CD3Z Plasma membrane 2.14803062 1.60E−11 polypeptide (TiT3 complex) 206914_at class-I MHC- CRTAM Plasma membrane 1.697660887 9.47E−10 restricted T cell associated molecule 205291_at interleukin 2 IL2RB Plasma membrane 1.700032107 1.14E−10 receptor, beta | interleukin 2 receptor, beta 204116_at interleukin 2 IL2RG Plasma membrane 2.261825374 4.95E−11 receptor, gamma (severe combined immunodeficiency) 206398_s_at CD19 antigen CD19 Plasma membrane 2.872617474 3.29E−16 217001_x_at major HLA-DOA Plasma membrane 1.589617654 1.39E−08 histocompatibility complex, class II, DO alpha 226878_at Major HLA-DOA Plasma membrane 1.561941309 3.01E−06 histocompatibility complex, class II, DO alpha 205671_s_at major HLA-DOB Plasma membrane 4.102368289 4.62E−17 histocompatibility complex, class II, DO beta 205049_s_at CD79A antigen CD79A Plasma membrane 6.214381039 7.29E−15 (immunoglobulin- associated alpha) | CD79A antigen (immunoglobulin- associated alpha) 1555779_a_at CD79A antigen CD79A Plasma membrane 3.486445366 1.47E−17 (immunoglobulin- associated alpha) 205297_s_at CD79B antigen CD79B Plasma membrane 2.131525118 6.42E−15 (immunoglobulin- associated beta) 224404_s_at immunoglobulin IRTA2 Plasma membrane 7.598702121 6.29E−20 superfamily receptor translocation associated 2 | immunoglobulin superfamily receptor translocation associated 2 224405_at immunoglobulin IRTA2 Plasma membrane 4.857349464 1.42E−19 superfamily receptor translocation associated 2 | immunoglobulin superfamily receptor translocation associated 2 224406_s_at immunoglobulin IRTA2 Plasma membrane 3.753764961 2.57E−20 superfamily receptor translocation associated 2 | immunoglobulin superfamily receptor translocation associated 2 231647_s_at immunoglobulin IRTA2 Plasma membrane 11.05597709 9.47E−21 superfamily receptor translocation associated 2 205692_s_at CD38 antigen (p45) CD38 Plasma membrane 3.802517688 9.27E−20 236191_at CD38 antigen (p45) CD38 Plasma membrane 2.235267342 6.23E−08 1552584_at interleukin 12 IL12RB1 Plasma membrane 1.551031389 2.43E−05 receptor, beta 1 206999_at interleukin 12 IL12RB2 Plasma membrane 1.75915602 1.04E−05 receptor, beta 2 219971_at interleukin 21 IL21R Plasma membrane 2.933140561 1.33E−09 receptor 221658_s_at interleukin 21 IL21R Plasma membrane 1.915118011 7.74E−08 receptor 205242_at chemokine (C-X-C CXCL13 Extracellular 5.900697331 4.68E−05 motif) ligand 13 (B- (Secreted) cell chemoattractant) 207339_s_at lymphotoxin beta LTB Plasma membrane 2.807618909 1.70E−09 (TNF superfamily, member 3) 204949_at intercellular adhesion ICAM3 Plasma membrane 2.650287454 3.47E−17 molecule 3 206295_at interleukin 18 IL18 Extracellular 1.607893049 2.43E−07 (interferon-gamma- (Secreted) inducing factor) 211633_x_at Immunoglobulin IGHG1 Extracellular 9.72344799 1.95E−13 heavy constant (Secreted) gamma 1 (G1m marker) | Immunoglobulin heavy constant gamma 1 (G1m marker) 209374_s_at immunoglobulin IGHM Extracellular 3.479239038 2.11E−12 heavy constant mu (Secreted) 221286_s_at proapoptotic caspase PACAP Extracellular 22.71584082 1.30E−15 adaptor protein (Secreted) 223565_at proapoptotic caspase PACAP Extracellular 14.28785156 4.53E−18 adaptor protein (Secreted)

Using the molecular phenotype training and testing statistical methods described above, an L phenotype classifier was identified, as indicated in Table 10 below.

TABLE 10 L phenotype (subtype) classifier genes and probes NETAFFX: ProbeID NETAFFX: GeneTitle GeneSymbol 234366_x_at 28396&3493&3500 NA (Entrez Gene Identifiers) 217235_x_at 28396&3493&3500 NA (Entrez Gene Identifiers) 236401_at 170575&26157 NA (Entrez Gene Identifiers) 200670_at Xbox binding protein 1 XBP1 1554050_at sphingomyelin phosphodiesterase SMPDL3B acidlike 3B 1556180_at hypothetical protein LOC255458 LOC255458 217480_x_at similar to Ig kappa chain LOC339562 204269_at pim2 oncogene PIM2 232167_at Solute carrier family 2 facilitated SLC2A11 glucose transporter member 11 226811_at family with sequence similarity 46 FAM46C member C 238560_at nuclear domain 10 protein NDP52 1555779_a_at CD79A antigen CD79A immunoglobulinassociated alpha 218237_s_at solute carrier family 38 member 1 SLC38A1 226773_at MRNA clone ICRFp507I1077 NA 215946_x_at immunoglobulin lambdalike IGLL1 & polypeptide 1 & similar to LOC91316 bK246H31 immunoglobulin lambdalike polypeptide 1 preBcell specific 204613_at phospholipase C gamma 2 PLCG2 phosphatidylinositolspecific 217369_at similar to immunoglobulin M chain LOC440361 216542_x_at hypothetical protein MGC27165 MGC27165 216207_x_at immunoglobulin kappa IGKV1D13 variable 1D13 201998_at ST6 betagalactosamide ST6GAL1 alpha26sialyltranferase 1 217168_s_at homocysteineinducible HERPUD1 endoplasmic reticulum stressinducible ubiquitinlike domain member 1 212204_at DKFZP564G2022 protein DKFZP564G2022 64064_at GTPase IMAP family member 5 GIMAP5 229721_x_at Der1like domain family member 3 DERL3 219435_at hypothetical protein FLJ22170 FLJ22170 203335_at phytanoylCoA hydroxylase Refsum PHYH disease 231611_at NA NA 214268_s_at myotubularin related protein 4 MTMR4 208093_s_at nudE nuclear distribution gene E NDEL1 homolog like 1 A nidulans & nudE nuclear distribution gene E homolog like 1 A nidulans 226075_at SPRY domaincontaining SOCS box SSB1 protein SSB1 213118_at NA NA 218318_s_at nemo like kinase NLK 227326_at Transmembrane anchor protein 1 TMAP1 223917_s_at solute carrier family 39 zinc SLC39A3 transporter member 3 208056_s_at corebinding factor runt domain alpha CBFA2T3 subunit 2; translocated to 3 231016_s_at NA NA 202916_s_at family with sequence similarity 20 FAM20B member B 217390_x_at NA NA 219118_at FK506 binding protein 11 19 kDa FKBP11 229686_at purinergic receptor P2Y Gprotein P2RY8 coupled 8 212311_at KIAA0746 protein KIAA0746 212699_at secretory carrier membrane protein 5 SCAMP5 219631_at low density lipoproteinrelated LRP12 protein 12 202089_s_at solute carrier family 39 zinc SLC39A6 transporter member 6 1555981_at hypothetical protein DKFZp762C2414 DKFZp762C2414 220647_s_at E2IG2 protein E2IG2 237383_at NA NA 230337_at son of sevenless homolog 1 SOS1 Drosophila 1559820_at APG10 autophagy 10like S cerevisiae APG10L

Another grouping of samples shown in FIG. 2 (21%) defined the molecular subtype described herein as myeloid-rich (M) (FIG. 2, the branches at the top of the dendrogram labeled “M” at the top of figure and corresponding box shown within the heatmap, 67% branch support). The gene expression signature of this group of samples revealed a pattern characteristic of monocytes, macrophages, neutrophils, and lymphocytes and implicated certain pathways including macrophage activation, phagocytosis, respiratory burst, T cell activation and cytokine production. In addition, this subtype was inversely associated with joint vascularity (p<0.05). Table 6 provides a list of probe sets (and associated genes) that are associated with the M subtype. These were identified from the microarray data using the following criteria: (1) Fold change≧1.5 within the M subtype; (2) t-test p-value≦0.0001; and (3) annotated as belonging to at least one of the following molecular categories of proteins: secreted, plasma membrane, kinase, G-coupled protein receptor, phosphatase, nuclear receptor, ion channel, E3-ligase, de-ubiquitinating enzyme.

Table 2 below shows a subset of certain of these probe sets (and associated genes) from Table 6 that have been identified as therapeutic targets and biomarkers of the M subtype. The genes identified in Table 2 encode proteins that share the properties of surface expression and secretion. Proteins having those properties can, in certain instances, be targeted with, for example, monoclonal antibodies and in that case are considered therapeutic targets. Secreted proteins and products cleaved from the cell membrane can, in certain cases, be measured and in that case are considered biomarkers.

TABLE 2 Certain M subtype therapeutic targets and biomarkers. NETAFFX: NETAFFX: Unison: Fold ProbeID GeneTitle GeneSymbol Protcomp Change p-value 219890_at C-type lectin domain CLEC5A Extracellular 7.362977 2.42E−07 family 5, member A (Secreted) 221698_s_at C-type lectin domain CLEC7A Plasma membrane 2.051059 5.18E−09 family 7, member A | C-type lectin domain family 7, member A 1555214_a_at C-type lectin domain CLEC7A Plasma membrane 2.246684 0.000198 family 7, member A 1554406_a_at C-type lectin domain CLEC7A Plasma membrane 1.581093 1.30E−07 family 7, member A 1555756_a_at C-type lectin domain CLEC7A Plasma membrane 1.893079 1.41E−06 family 7, member A 201951_at Activated leukocyte ALCAM Plasma membrane 2.174859 7.52E−10 cell adhesion molecule 210233_at interleukin 1 receptor IL1RAP Plasma membrane 2.13583 8.45E−10 accessory protein 201587_s_at interleukin-1 receptor- IRAK1 Nuclear 1.523179 3.11E−09 associated kinase 1 210842_at neuropilin 2 NRP2 Plasma membrane 3.63583 1.52E−05 232701_at Neuropilin 2 NRP2 Plasma membrane 1.892335 2.23E−05 219434_at triggering receptor TREM1 Extracellular 2.054149 2.84E−08 expressed on myeloid (Secreted) cells 1 210512_s_at vascular endothelial VEGF Extracellular 2.563299 4.62E−11 growth factor (Secreted) 210513_s_at vascular endothelial VEGF Extracellular 1.635665 1.97E−09 growth factor (Secreted) 211527_x_at vascular endothelial VEGF Extracellular 3.030532 2.35E−13 growth factor (Secreted) 212171_x_at vascular endothelial VEGF Extracellular 2.323697 2.23E−14 growth factor (Secreted) 205179_s_at a disintegrin and ADAM8 Plasma membrane 2.429618 3.40E−06 metalloproteinase domain 8 | a disintegrin and metalloproteinase domain 8 205180_s_at a disintegrin and ADAM8 Plasma membrane 2.620813 1.03E−07 metalloproteinase domain 8 | a disintegrin and metalloproteinase domain 8 213274_s_at cathepsin B CTSB Lysosomal 1.656338 2.02E−09 207850_at chemokine (C-X-C CXCL3 Extracellular 2.692214 2.61E−07 motif) ligand 3 (Secreted) 202637_s_at intercellular adhesion ICAM1 Plasma membrane 2.12742 8.01E−14 molecule 1 (CD54), human rhinovirus receptor 202638_s_at intercellular adhesion ICAM1 Plasma membrane 2.297289 1.58E−17 molecule 1 (CD54), human rhinovirus receptor 215485_s_at intercellular adhesion ICAM1 Plasma membrane 2.070317 1.48E−12 molecule 1 (CD54), human rhinovirus receptor 222868_s_at interleukin 18 binding IL18BP Extracellular 1.523947 5.91E−06 protein (Secreted) 205067_at interleukin 1, beta IL1B Extracellular 1.680015 0.000269 (Secreted) 39402_at interleukin 1, beta IL1B Extracellular 2.550957 2.81E−06 (Secreted) 202859_x_at interleukin 8 IL8 Extracellular 3.188671 1.62E−05 (Secreted) 211506_s_at interleukin 8 IL8 Extracellular 4.223337 1.08E−06 (Secreted) 204580_at matrix MMP12 Extracellular 8.298394 5.23E−07 metalloproteinase 12 (Secreted) (macrophage elastase) 200660_at S100 calcium binding S100A11 NO_LOCALIZATION 1.574125 3.46E−11 protein A11 (calgizzarin)

Using the molecular phenotype training and testing statistical methods described above, an M phenotype classifier was identified, as indicated in Table 11 below.

TABLE 11 M phenotype (subtype) classifier genes and probes NETAFFX: ProbeID NETAFFX: GeneTitle GeneSymbol 224374_s_at elastin microfibril interfacer 2 & elastin microfibril interfacer 2 EMILIN2 203175_at ras homolog gene family member G rho G RHOG 212715_s_at flavoprotein oxidoreductase MICAL3 MICAL3 202944_at Nacetylgalactosaminidase alpha NAGA 212268_at serine or cysteine proteinase inhibitor clade B ovalbumin member 1 SERPINB1 202638_s_at intercellular adhesion molecule 1 CD54 human rhinovirus receptor ICAM1 200808_s_at zyxin ZYX 226587_at Ubiquitin protein ligase E3A human papilloma virus UBE3A E6associated protein Angelman syndrome 214785_at vacuolar protein sorting 13A yeast VPS13A 218627_at hypothetical protein FLJ11259 FLJ11259 216598_s_at chemokine CC motif ligand 2 CCL2 1557915_s_at glutathione Stransferase omega 1 GSTO1 207697_x_at leukocyte immunoglobulinlike receptor subfamily B with TM and LILRB2 ITIM domains member 2 214752_x_at filamin A alpha actin binding protein 280 FLNA 210225_x_at leukocyte immunoglobulinlike receptor subfamily B with TM and LILRB3 ITIM domains member 3 & leukocyte immunoglobulinlike receptor subfamily B with TM and ITIM domains member 3 204088_at purinergic receptor P2X ligandgated ion channel 4 P2RX4 211133_x_at leukocyte immunoglobulinlike receptor subfamily B with TM and LILRB3 ITIM domains member 3 227026_at Mphase phosphoprotein mpp8 HSMPP8 208018_s_at hemopoietic cell kinase HCK 208691_at NA NA 207850_at chemokine CXC motif ligand 3 CXCL3 210784_x_at leukocyte immunoglobulinlike receptor subfamily B with TM and LILRB3 ITIM domains member 3 209344_at tropomyosin 4 TPM4 201950_x_at capping protein actin filament muscle Zline beta CAPZB 201118_at phosphogluconate dehydrogenase & phosphogluconate PGD dehydrogenase 227961_at Cathepsin B CTSB 212041_at ATPase H+ transporting lysosomal 38 kDa V0 subunit d isoform 1 ATP6V0D1 202856_s_at solute carrier family 16 monocarboxylic acid transporters member 3 SLC16A3 210042_s_at cathepsin Z CTSZ 220088_at complement component 5 receptor 1 C5a ligand C5R1 219053_s_at Hypothetical protein FLJ20847 FLJ20847 214683_s_at CDClike kinase 1 CLK1 212171_x_at vascular endothelial growth factor VEGF 223019_at chromosome 9 open reading frame 88 C9orf88 212481_s_at tropomyosin 4 TPM4 202206_at ADPribosylation factorlike 7 ARL7 226389_s_at Rap guanine nucleotide exchange factor GEF 1 RAPGEF1 207332_s_at transferrin receptor p90 CD71 TFRC 202637_s_at intercellular adhesion molecule 1 CD54 human rhinovirus receptor ICAM1 201972_at ATPase H+ transporting lysosomal 70 kDa V1 subunit A ATP6V1A 215706_x_at zyxin ZYX 222877_at Neuropilin 2 NRP2 205098_at chemokine CC motif receptor 1 CCR1 202679_at NiemannPick disease type C1 NPC1 241684_at Transcribed locus weakly similar to NP_0603121 NA hypothetical protein FLJ20489 [Homo sapiens] 216035_x_at transcription factor 7like 2 Tcell specific HMGbox TCF7L2 242824_at KIAA0485 protein KIAA0485 202207_at ADPribosylation factorlike 7 ARL7 204137_at transmembrane 7 superfamily member 1 upregulated in kidney TM7SF1 205479_s_at plasminogen activator urokinase PLAU 202087_s_at cathepsin L CTSL 218665_at frizzled homolog 4 Drosophila FZD4 226354_at lactamase beta LACTB 212675_s_at KIAA0582 KIAA0582 213457_at malignant fibrous histiocytoma amplified sequence 1 MFHAS1 211135_x_at leukocyte immunoglobulinlike receptor subfamily B with TM and LILRB3 ITIM domains member 3 213313_at RAB GTPase activating protein 1 RABGAP1 215485_s_at intercellular adhesion molecule 1 CD54 human rhinovirus ICAM1 receptor 205775_at family with sequence similarity 50 member B FAM50B 210512_s_at vascular endothelial growth factor VEGF 202484_s_at methylCpG binding domain protein 2 MBD2 1560060_s_at vacuolar protein sorting 37C yeast VPS37C 211160_x_at actinin alpha 1 ACTN1 230528_s_at hypothetical protein MGC2752 MGC2752 210845_s_at plasminogen activator urokinase receptor PLAUR 213571_s_at eukaryotic translation initiation factor 4E member 2 EIF4E2

As shown in FIG. 2, two non-inflammatory molecular subtypes were identified. The first grouping (22%) defined the molecular subtype described herein as fibroblast-rich type 2 (F2) (FIG. 2, the branches at the top of the dendrogram labeled “F2” at the top of figure and corresponding box shown within the heatmap, 94% branch support). The gene expression signature of this group of samples revealed a pattern characteristic of fibroblasts and osteoblasts and implicated certain pathways including bone formation, growth and differentiation, wnt-signaling and tumorigenesis. The F2 gene expression signature was inversely associated with lymphocyte and CD15+ cell infiltration (each p<0.01). The second non-inflammatory grouping (11%) defined the molecular subtype described herein as fibroblast-rich type 1 (F1) (FIG. 2, the branches at the top of the dendrogram labeled “F1” at the top of figure and corresponding box shown within the heatmap, 88% branch support). The gene expression signature of this group of samples revealed a pattern characteristic of fibroblasts, osteoclasts, and osteoblasts and implicated certain pathways including bone destruction and vasculogenesis. In addition, the F1 subtype was associated with a higher degree of synovial lining hyperplasia compared to the other subtypes. Tables 7 and 8 provide a list of probe sets (and associated genes) that are associated with F2 and F1 subtypes, respectively. These were identified from the microarray data using the following criteria: (1) Fold change≧1.5 within the L subtype; (2) t-test p-value≦0.0001; and (3) annotated as belonging to at least one of the following molecular categories of proteins: secreted, plasma membrane, kinase, G-coupled protein receptor, phosphatase, nuclear receptor, ion channel, E3-ligase, de-ubiquitinating enzyme.

Table 3 below shows a subset of certain of these probe sets (and associated genes) from Table 7 that have been identified as therapeutic targets and biomarkers of the F2 subtype. Table 4 below shows a subset of certain of these probe sets (and associated genes) from Table 8 that have been identified as therapeutic targets and biomarkers of the F1 subtype. The genes identified in Tables 3 and 4 encode proteins that share the properties of surface expression and secretion. Proteins having those properties can, in certain instances, be targeted with, for example, monoclonal antibodies and in that case are considered therapeutic targets. Secreted proteins and products cleaved from the cell membrane can, in certain cases, be measured and in that case are considered biomarkers.

TABLE 3 Certain F2 subtype therapeutic targets and biomarkers. NETAFFX: NETAFFX: Unison: Fold ProbeID GeneTitle GeneSymbol Protcomp Change p-value 227401_at interleukin 17D IL17D Extracellular 2.453105 2.26E−12 (Secreted) 228977_at Interleukin 17D IL17D NA 1.999348 1.75E−11 221926_s_at interleukin 17 IL17RC NO_LOCALIZATION 2.406789 8.12E−09 receptor C 201147_s_at tissue inhibitor of TIMP3 Extracellular 1.872803 9.91E−13 metalloproteinase 3 (Secreted) (Sorsby fundus dystrophy, pseudoinflammatory) 201148_s_at tissue inhibitor of TIMP3 Extracellular 1.915979 4.22E−12 metalloproteinase 3 (Secreted) (Sorsby fundus dystrophy, pseudoinflammatory) 201149_s_at tissue inhibitor of TIMP3 Extracellular 2.064168 1.45E−11 metalloproteinase 3 (Secreted) (Sorsby fundus dystrophy, pseudoinflammatory) 240135_x_at Tissue inhibitor of TIMP3 Extracellular 2.474079 4.55E−07 metalloproteinase 3 (Secreted) (Sorsby fundus dystrophy, pseudoinflammatory) 204932_at tumor necrosis factor TNFRSF11B Extracellular 3.094446 1.47E−07 receptor superfamily, (Secreted) member 11b (osteoprotegerin) 204933_s_at tumor necrosis factor TNFRSF11B Extracellular 3.791135 8.36E−08 receptor superfamily, (Secreted) member 11b (osteoprotegerin) 231762_at fibroblast growth factor FGF10 Extracellular 2.530553 4.25E−06 10 (Secreted) 206987_x_at fibroblast growth factor FGF18 Extracellular 2.36539 1.98E−05 18 (Secreted) 211029_x_at fibroblast growth factor FGF18 Extracellular 2.332917 2.81E−05 18 | fibroblast growth (Secreted) factor 18 211485_s_at fibroblast growth factor FGF18 Extracellular 2.955372 1.03E−05 18 (Secreted) 231382_at Fibroblast growth FGF18 Extracellular 2.766924 1.55E−07 factor 18 (Secreted) 204421_s_at fibroblast growth factor FGF2 NO_LOCALIZATION 1.56491 3.28E−07 2 (basic) 204422_s_at fibroblast growth factor FGF2 NO_LOCALIZATION 2.144299 1.38E−07 2 (basic) 205606_at low density lipoprotein LRP6 Plasma membrane 1.935109 4.17E−05 receptor-related protein 6 209909_s_at transforming growth TGFB2 Extracellular 2.066203 4.94E−15 factor, beta 2 (Secreted) 220407_s_at transforming growth TGFB2 Extracellular 1.711286 8.48E−10 factor, beta 2 (Secreted) 228121_at Transforming growth TGFB2 Extracellular 1.699841 3.07E−13 factor, beta 2 (Secreted) 206737_at wingless-type MMTV WNT11 Extracellular 3.235511 3.83E−07 integration site family, (Secreted) member 11 206176_at bone morphogenetic BMP6 Extracellular 1.617235 2.14E−06 protein 6 (Secreted) 207326_at betacellulin BTC Plasma membrane 4.484959 6.59E−11 241412_at betacellulin BTC Plasma membrane 5.748075 1.57E−12 219764_at frizzled homolog 10 FZD10 Plasma membrane 1.886755 9.08E−10 (Drosophila) 203705_s_at frizzled homolog 7 FZD7 Plasma membrane 1.977168 1.12E−13 (Drosophila) 203706_s_at frizzled homolog 7 FZD7 Plasma membrane 2.148229 7.07E−15 (Drosophila) 224325_at frizzled homolog 8 FZD8 Plasma membrane 2.039929 2.47E−13 (Drosophila) | frizzled homolog 8 (Drosophila) 227405_s_at frizzled homolog 8 FZD8 Plasma membrane 2.033817 1.11E−13 (Drosophila)

TABLE 4 Certain F1 subtype therapeutic targets and biomarkers. NETAFFX: NETAFFX: Unison: Fold ProbeID GeneTitle GeneSymbol Protcomp Change p-value 236179_at Cadherin 11, type 2, CDH11 Plasma membrane 2.478118072 1.48E−13 OB-cadherin (osteoblast) 239286_at Cadherin 11, type 2, CDH11 Plasma membrane 2.003613835 1.78E−09 OB-cadherin (osteoblast) 241780_at Cadherin 11, type 2, CDH11 Plasma membrane 1.991499524 5.56E−06 OB-cadherin (osteoblast) 207173_x_at cadherin 11, type 2, CDH11 Plasma membrane 1.848606565 1.15E−11 OB-cadherin (osteoblast) 207172_s_at cadherin 11, type 2, CDH11 Plasma membrane 1.803978278 1.00E−10 OB-cadherin (osteoblast) 222899_at integrin, alpha 11 ITGA11 Plasma membrane 2.22681306 1.32E−08 205131_x_at C-type lectin domain CLEC11A Cytoplasmic 2.751077434 1.15E−12 family 11, member A 211709_s_at C-type lectin domain CLEC11A Cytoplasmic 2.09446091 3.80E−11 family 11, member A | C-type lectin domain family 11, member A 210783_x_at C-type lectin domain CLEC11A Cytoplasmic 2.078646468 3.56E−06 family 11, member A 203876_s_at matrix MMP11 Plasma membrane 2.200173425 8.85E−06 metalloproteinase 11 (stromelysin 3) 235908_at matrix MMP11 Plasma membrane 1.952831042 1.58E−10 metalloproteinase 11 (stromelysin 3) 205959_at matrix MMP13 Extracellular 8.887243061 2.09E−09 metalloproteinase (Secreted) 13 (collagenase 3) | matrix metalloproteinase 13 (collagenase 3) 207012_at matrix MMP16 Plasma membrane 1.749078693 2.81E−08 metalloproteinase 16 (membrane- inserted) 224207_x_at matrix MMP28 Extracellular 2.533723399 3.21E−09 metalloproteinase (Secreted) 28 222937_s_at matrix MMP28 Extracellular 1.616679576 9.53E−07 metalloproteinase (Secreted) 28 213790_at A disintegrin and ADAM12 Plasma membrane 6.765178528 3.70E−12 metalloproteinase domain 12 (meltrin alpha) 226777_at A disintegrin and ADAM12 Plasma membrane 5.054380937 1.32E−12 metalloproteinase domain 12 (meltrin alpha) 202952_s_at a disintegrin and ADAM12 Plasma membrane 4.582308224 1.37E−11 metalloproteinase domain 12 (meltrin alpha) 241026_at A disintegrin and ADAM12 Plasma membrane 3.111313526 1.10E−05 metalloproteinase domain 12 (meltrin alpha) 215613_at A disintegrin and ADAM12 Plasma membrane 2.348274812 3.16E−07 metalloproteinase domain 12 (meltrin alpha) 208227_x_at a disintegrin and ADAM22 Plasma membrane 3.465935261 1.11E−09 metalloproteinase domain 22 202450_s_at cathepsin K CTSK Lysosomal 1.746776635 4.32E−14 (pycnodysostosis) 211148_s_at angiopoietin 2 ANGPT2 Extracellular 1.961743434 3.09E−10 (Secreted) 205572_at angiopoietin 2 ANGPT2 Extracellular 1.858241241 5.95E−08 (Secreted) 236034_at Angiopoietin 2 ANGPT2 Extracellular 1.74308462 8.68E−08 (Secreted) 223121_s_at secreted frizzled- SFRP2 Extracellular 3.420031277 5.12E−07 related protein 2 (Secreted) 223122_s_at secreted frizzled- SFRP2 Extracellular 2.45294325 4.10E−07 related protein 2 (Secreted) 204051_s_at secreted frizzled- SFRP4 Extracellular 2.115342409 8.29E−05 related protein 4 (Secreted) 204468_s_at tyrosine kinase with TIE1 Plasma membrane 2.020004455 2.91E−07 immunoglobulin-like and EGF-like domains 1 202112_at von Willebrand VWF Extracellular 1.858748797 1.03E−08 factor (Secreted)

Using the molecular phenotype training and testing statistical methods described above, an F2 phenotype classifier was identified, as indicated in Table 12 below.

TABLE 12 F2 phenotype (subtype) classifier genes and probes NETAFFX: ProbeID NETAFFX: GeneTitle GeneSymbol 201802_at solute carrier family 29 nucleoside SLC29A1 transporters member 1 227405_s_at frizzled homolog 8 Drosophila FZD8 204237_at GULP engulfment adaptor GULP1 PTB domain containing 1 205158_at ribonuclease RNase A family 4 RNASE4 227526_at Full length insert cDNA clone NA ZD42A08 204235_s_at GULP engulfment adaptor PTB GULP1 domain containing 1 203554_x_at pituitary tumortransforming 1 PTTG1 201801_s_at solute carrier family 29 nucleoside SLC29A1 transporters member 1 206002_at G proteincoupled receptor 64 GPR64 212914_at chromobox homolog 7 CBX7 228084_at Fulllength cDNA clone NA CS0DF027YF17 of Fetal brain of Homo sapiens human 222043_at clusterin complement lysis inhibitor CLU SP4040 sulfated glycoprotein 2 testosteronerepressed prostate message 2 apolipoprotein J 229310_at kelch repeat and BTB POZ domain KBTBD9 containing 9 225728_at importin 9 IPO9 226247_at pleckstrin homology domain PLEKHA1 containing family A phosphoinositide binding specific member 1 205794_s_at neurooncological ventral antigen 1 NOVA1 213497_at ankyrin repeat and BTB POZ domain ABTB2 containing 2 207551_s_at malespecific lethal 3like 1 Drosophila MSL3L1 227554_at Hypothetical LOC402560 NA 223315_at netrin 4 NTN4 208868_s_at GABAA receptorassociated protein GABARAPL1 like 1 210046_s_at isocitrate dehydrogenase 2 NADP+ IDH2 mitochondrial 219295_s_at procollagen Cendopeptidase enhancer 2 PCOLCE2 221796_at neurotrophic tyrosine kinase receptor NTRK2 type 2 208869_s_at GABAA receptorassociated protein GABARAPL1 like 1 225950_at Transcribed locus moderately similar to NA NP_0553011 neuronal thread protein AD7cNTP [Homo sapiens] 204288_s_at ArgAblinteracting protein ArgBP2 ARGBP2 223842_s_at scavenger receptor class A member 3 SCARA3 203306_s_at solute carrier family 35 CMPsialic acid SLC35A1 transporter member A1 208792_s_at clusterin complement lysis inhibitor CLU SP4040 sulfated glycoprotein 2 testosteronerepressed prostate message 2 apolipoprotein J 203744_at highmobility group box 3 HMGB3 1555778_a_at periostin osteoblast specific factor POSTN 227308_x_at latent transforming growth factor beta LTBP3 binding protein 3 229969_at Transcribed locus weakly similar to NA NP_0790122 gasdermin domain containing 1 [Homo sapiens] 224989_at Hypothetical protein LOC201895 LOC201895 222423_at Nedd4 family interacting protein 1 NDFIP1 227052_at Hypothetical protein LOC201895 LOC201895 230351_at hypothetical protein LOC283481 LOC283481 219230_at hypothetical protein FLJ10970 FLJ10970 226197_at Transcribed locus strongly similar to NA XP_4960551 similar to p40 [Homo sapiens] 212599_at autism susceptibility candidate 2 AUTS2 203805_s_at Fanconi anemia complementation FANCA group A & Fanconi anemia complementation group A 202429_s_at protein phosphatase 3 formerly 2B PPP3CA catalytic subunit alpha isoform calcineurin A alpha 218471_s_at BardetBiedl syndrome 1 BBS1 227290_at CDNA FLJ13598 fis clone NA PLACE1009921 1552790_a_at hypothetical protein FLJ32803 FLJ32803 212616_at chromodomain helicase DNA binding CHD9 protein 9

Using the molecular phenotype training and testing statistical methods described above, an F1 phenotype classifier was identified, as indicated in Table 13 below.

TABLE 13 F1 phenotype (subtype) classifier genes and probes NETAFFX: ProbeID NETAFFX: GeneTitle GeneSymbol 213059_at cAMP responsive element binding protein 3like 1 CREB3L1 204385_at kynureninase Lkynurenine hydrolase KYNU 207172_s_at cadherin 11 type 2 OBcadherin osteoblast CDH11 212771_at chromosome 10 open reading frame 38 C10orf38 218454_at hypothetical protein FLJ22662 FLJ22662 203010_at signal transducer and activator of transcription 5A STAT5A 210992_x_at Fc fragment of IgG low affinity IIc receptor for CD32 FCGR2C 203903_s_at hephaestin HEPH 227307_at Tetraspanin similiar to uroplakin 1 LOC90139 201307_at septin 11 SEPT11 202902_s_at cathepsin S CTSS 202897_at protein tyrosine phosphatase nonreceptor type substrate 1 PTPNS1 226098_at KIAA1374 protein KIAA1374 206116_s_at tropomyosin 1 alpha TPM1 204787_at Vset and immunoglobulin domain containing 4 VSIG4 230264_s_at adaptorrelated protein complex 1 sigma 2 subunit AP1S2 227618_at FLJ44635 protein FLJ44635 206571_s_at mitogenactivated protein kinase kinase kinase kinase 4 MAP4K4 217984_at ribonuclease T2 RNASET2 212276_at lipin 1 LPIN1 203417_at microfibrillarassociated protein 2 MFAP2 223614_at NA NA 211981_at collagen type IV alpha 1 COL4A1 226828_s_at hairyenhancerofsplit related with YRPW motiflike HEYL 209081_s_at collagen type XVIII alpha 1 COL18A1 228396_at NA NA 224759_s_at hypothetical protein MGC17943 MGC17943 212951_at G proteincoupled receptor 116 GPR116 222664_at potassium channel tetramerisation domain containing 15 KCTD15 221942_s_at guanylate cyclase 1 soluble alpha 3 GUCY1A3 205819_at macrophage receptor with collagenous structure & MARCO macrophage receptor with collagenous structure 204677_at cadherin 5 type 2 VEcadherin vascular epithelium CDH5 228339_at Transcribed locus strongly similar to XP_5308421 NA LOC462106 [Pan troglodytes] 227235_at Homo sapiens clone IMAGE: 5302158 mRNA NA 219489_s_at nucleoredoxin NXN 203299_s_at adaptorrelated protein complex 1 sigma 2 subunit AP1S2 203300_x_at adaptorrelated protein complex 1 sigma 2 subunit AP1S2 224749_at chromosome 16 open reading frame 9 C16orf9 210663_s_at kynureninase Lkynurenine hydrolase KYNU 227333_at Hypothetical protein MGC48972 MGC48972 209696_at fructose16bisphosphatase 1 FBP1 212985_at Fulllength cDNA clone CS0DC015YK09 of NA Neuroblastoma Cot 25normalized of Homo sapiens human 226575_at zinc finger protein 462 ZNF462 229121_at CDNA FLJ44441 fis clone UTERU2020242 NA 214770_at macrophage scavenger receptor 1 MSR1 201401_s_at adrenergic beta receptor kinase 1 ADRBK1 227627_at serumglucocorticoid regulated kinaselike SGKL 218041_x_at solute carrier family 38 member 2 SLC38A2 217846_at glutaminyltRNA synthetase QARS 203507_at CD68 antigen CD68 203505_at ATPbinding cassette subfamily A ABC1 member 1 ABCA1 202418_at Yip1 interacting factor homolog S cerevisiae YIF1 221685_s_at hypothetical protein FLJ20364 FLJ20364 230422_at formyl peptide receptorlike 2 FPRL2 226084_at microtubuleassociated protein 1B MAP1B 203923_s_at cytochrome b245 beta polypeptide chronic CYBB granulomatous disease 211208_s_at calciumcalmodulindependent serine protein kinase CASK MAGUK family 219694_at hypothetical protein FLJ11127 FLJ11127 217388_s_at kynureninase Lkynurenine hydrolase KYNU 1555778_a_at periostin osteoblast specific factor POSTN 1554285_at hepatitis A virus cellular receptor 2 HAVCR2 204834_at fibrinogenlike 2 FGL2 203148_s_at tripartite motifcontaining 14 TRIM14 207857_at leukocyte immunoglobulinlike receptor subfamily A LILRA2 with TM domain member 2 & leukocyte immunoglobulinlike receptor subfamily A with TM domain member 2 238668_at Transcribed locus NA 232617_at cathepsin S CTSS 217983_s_at ribonuclease T2 RNASET2 1555349_a_at integrin beta 2 antigen CD18 p95 lymphocyte ITGB2 functionassociated antigen 1; macrophage antigen 1 mac1 beta subunit 211395_x_at Fc fragment of IgG low affinity IIc receptor for CD32 FCGR2C 204006_s_at Fc fragment of IgG low affinity IIIa receptor CD16a & FCGR3A & Fc fragment of IgG low affinity IIIb receptor CD16b FCGR3B 210629_x_at leukocyte specific transcript 1 LST1 218961_s_at polynucleotide kinase 3phosphatase PNKP 203922_s_at cytochrome b245 beta polypeptide chronic CYBB granulomatous disease 202803_s_at integrin beta 2 antigen CD18 p95 lymphocyte ITGB2 functionassociated antigen 1; macrophage antigen 1 mac1 beta subunit 209083_at coronin actin binding protein 1A CORO1A

To further characterize each of the molecular subtypes and find associations between the gene expression signature of each molecular subtype and clinical and histological features of RA, samples of the each of the molecular subtypes were analyzed for expression of one or more particular genes predominantly expressed in that subtype. Certain samples were also assessed for associations with systemic measure of inflammation, the erythroid sedimentation rate (ESR) and C-reactive protein (CRP) levels. Also, associations with radiographic progression were assessed. In addition, the samples were subjected to histological and immunohistochemical analyses.

FIG. 3 shows the results of these studies for the L subtype samples. FIG. 3A shows that the transcription factor XBP1 is upregulated in the L subtype samples (L) compared to samples of other subtypes (NL). Accordingly, expression of XBP1 is an L subtype-specific surrogate marker. Furthermore, FIG. 3B shows that XBP1 expression is significantly upregulated in synovial samples containing lymphoid aggregates (+) compared to samples lacking lymphoid aggregates (−). Box and whisker plots for FIGS. 3A and 3B represent each sample as an open circle. The box represents the 25th to 75th percentile and contains the median value (horizontal line within the box). The whiskers extend from the box to represent values up to 1.5 times above and below the interquartile range. FIG. 3 also shows that XBP1 expression is not associated with ESR (FIG. 3C) or CRP levels (FIG. 3D). FIGS. 3E-H show the results of histological and immunohistochemical staining of representative samples of the L subtype. FIG. 3E shows staining with hematoxylin and eosin; FIG. 3F shows immunohistochemical staining for the T cell marker CD3; FIG. 3G shows immunohistochemical staining for the activated leukocyte marker CD68; FIG. 3H shows immunohistochemical staining for the B cell marker CD20.

FIG. 4 shows the characterization of the M subtype samples. FIG. 4A shows that the gene ICAM1 is upregulated in the M subtype samples (M) compared to samples of the other subtypes (F1, F2, and L). In FIG. 4A, a box and whisker plot represents each sample as an open circle. The box represents the 25th to 75th percentile and contains the median value (horizontal line within the box). The whiskers extend from the box to represent values up to 1.5 times above and below the interquartile range. Accordingly, expression of ICAM1 is an M subtype-specific surrogate marker. FIG. 4B is a graphical plot of IL1β gene expression compared to TNF gene expression in M subtype samples. The plot shows that IL1β gene expression and TNF gene expression in synovial samples of the M subtype are correlated. This correlation was not observed in the other three molecular subtypes (data not shown). FIGS. 4C-F show the results of histological and immunohistochemical staining of representative samples of the M subtype. FIG. 4C shows staining with hematoxylin and eosin; FIG. 4D shows immunohistochemical staining for the T cell marker CD3; FIG. 4E shows immunohistochemical staining for the activated leukocyte marker CD68; FIG. 4F shows immunohistochemical staining for the B cell marker CD20.

FIG. 5 shows the characterization of the F2 subtype samples. FIG. 5A shows that the gene IL17D is upregulated in the F2 subtype samples (M) compared to samples of the other subtypes (F1, L, and M). In FIG. 5A, a box and whisker plot represents each sample as an open circle. The box represents the 25th to 75th percentile and contains the median value (horizontal line within the box). The whiskers extend from the box to represent values up to 1.5 times above and below the interquartile range. Accordingly, expression of IL17D is an F2 subtype-specific surrogate marker. FIGS. 5B-E show the results of histological and immunohistochemical staining of representative samples of the F2 subtype. FIG. 5B shows staining with hematoxylin and eosin; FIG. 5C shows immunohistochemical staining for the T cell marker CD3; FIG. 5D shows immunohistochemical staining for the activated leukocyte marker CD68; FIG. 5E shows immunohistochemical staining for the B cell marker CD20.

FIG. 6 shows the characterization of the F1 subtype samples. FIG. 6A shows that the gene ITGA11 is upregulated in the F1 subtype samples (M) compared to samples of the other subtypes (F2, L, and M). In FIG. 6A, a box and whisker plot represents each sample as an open circle. The box represents the 25th to 75th percentile and contains the median value (horizontal line within the box). The whiskers extend from the box to represent values up to 1.5 times above and below the interquartile range. Accordingly, expression of ITGA11 is an F1 subtype-specific surrogate marker. FIGS. 6B-E show the results of histological and immunohistochemical staining of representative samples of the F1 subtype. FIG. 6B shows staining with hematoxylin and eosin; FIG. 6C shows immunohistochemical staining for the T cell marker CD3; FIG. 6D shows immunohistochemical staining for the activated leukocyte marker CD68; FIG. 6E shows immunohistochemical staining for the B cell marker CD20.

For each of the subtypes, we determined the number of samples that were obtained from particular joints. This data is presented in Table 9 below.

TABLE 9 Distribution of joints by molecular subtype. Molecular Subtype Joint F1 F2 L M Elbow 0 0 0 1 Foot 0 1 0 0 Hand 0 2 4 10 Hip 2 1 2 0 Knee 1 4 5 1 Wrist 1 1 1 0

As indicated above, we observed follicle-like lymphoid clusters in the L subtype. We also analyzed histological sections of samples from each of the other three subtypes in addition to the L subtype and quantitated the percentage of samples within each subtype showing lymphoid clusters (or aggregates). The results are shown in FIG. 7. As shown in FIG. 7, approximately 60% of the L subtype samples had lymphoid clusters, whereas a much smaller percentage (<10%) of the F2 and M subtype samples had lymphoid clusters. An even smaller percentage of the F1 subtype samples (2%-3%) had lymphoid clusters. These results indicate that the L-subtype gene expression signature is associated with the presence of organized lymphoid structures within the joint.

The associations of each of the subtypes with systemic measures of inflammation, the erythroid sedimentation rate (ESR) and C-reactive protein (CRP) levels, were assessed, as well as associations of each of the subtypes with radiographic progression. ESR, CRP, and radiographic assessments were performed according to standard procedures well known to those skilled in the art. These associations are shown graphically in FIGS. 8A-C. As shown in FIGS. 8A-C, none of the subtypes is clearly associated with ESR, CRP and/or radiographic progression. As discussed above, ESR, CRP levels and radiographic progression have been employed as diagnostic markers in RA, each having certain limitations. See also Pinals, R. S., et. al., Arthritis Rheum 24:1308 (1981) and Felson, D. T., et al., Arthritis Rheum 38: 727-35 (1995). Accordingly, the gene expression signatures described here provide, for example but not limited to, new diagnostic compositions which can be employed using methods as described herein and which augment or circumvent limitations of prior diagnostic markers or methods.

To identify biological pathways implicated in each of the molecular subtypes, statistical analysis (pathway analysis) of the gene signatures specific to each subtype was performed. The results of this analysis are depicted in the heatmap shown in FIG. 9. Each molecular subtype is listed at the top of the heatmap and biological pathways are provided along the right side of the heatmap. The heatmap is shaded to represent the −log of the p-values for statistically enriched pathways within each subtype according to the scale shown at the bottom of the figure. Statistically enriched pathways were identified using a publicly available web-service, CoPub, following the developers' recommended protocol (Frijters, R. et al., Nucleic Acids Res. 36:W406-W410 (Web server issue doi:10.1093/nar/gkn215) (2008)); available at the URL: services(dot)nbic(dot)nl(slash)cgi-bin(slash)copub(slash)microarray_analysis(dot)pl. Briefly, Affymetrix probeset identifiers that were specifically upregulated within each subtype (˜1000 top ranked probesets) were uploaded to the web-server. The GeneChip® Human Genome U133A Plus 2.0 Array (Affymetrix, Inc.) was selected as the background data set, the search category was limited to biological processes and all calculation settings were left at their defaults. The resulting data was saved to a personal computer and formatted for comparative visualization. As indicated in FIG. 9, the biological pathways showing the highest statistical enrichment in the L subtype include, for example, B and T cell activation and cytokine production; the biological pathways showing the highest statistical enrichment in the M subtype include, for example, macrophage activation, phagocytosis, respiratory burst, and cytokine production; the biological pathways showing the highest statistical enrichment in the F2 subtype include, for example, bone formation, growth and differentiation, wnt-signaling and cell cycle; and the biological pathways showing the highest statistical enrichment in the F1 subtype include, for example, osteoblast differentiation, bone remodeling and vasculogenesis.

Example 2

To further characterize the molecular four phenotypes (subtypes) identified in Example 1, select genes representing the specific cellularities and biological processes of each phenotype were tested for specificity using real-time quantitative polymerase chain reaction (qPCR). As non-RA controls we used a set of synovial samples obtained from osteoarthritis patients (OA) and a set of synovial samples obtained from patients suffering from joint trauma but not from RA (Normal [Nrml]). Real-time qPCR was carried out as follows.

cDNA synthesis was performed using the iScript™ cDNA synthesis kit and protocol (Biorad, Hercules, Calif.). Two hundred ng of total RNA was added to a 20 μl cDNA reaction mixture containing 4 μl 5× iScript™ reaction mixture, 1 μl iScript™ reverse transcriptase and nuclease-free water. The reverse transcription reaction mixture was incubated at 25° C. for 5 minutes, 42° C. for 30 minutes and 85° C. for 5 minutes.

A gene specific pre-amplification of cDNA samples was performed using the TaqMan® PreAmp Master Mix (Applied Biosystems, Foster City, Calif.). One μl of a total of 77 20× TaqMan® Gene Expression Assays (all assays contained FAM™ dye-labeled MGB probes, Applied Biosystems, Foster City, Calif.) were pooled and diluted with 1×TE buffer for a final concentration of 0.2× per assay. Per sample, 1.25 μl of cDNA, 1.25 μl of the pooled assay mix and 2.5 μl of 2× TaqMan® PreAmp Master Mix (Applied Biosystems) were mixed. The pre-amplification reactions were done in a GeneAmp® PCR System 9700 (Applied Biosystems, Foster City, Calif.) using the protocol, 95° C. for 10 minutes, and 14 cycles of 95° C. for 15 seconds and 60° C. for 4 minutes. After thermal cycling, the pre-amplified samples were diluted five times with 1×TE buffer.

Semi-quantitative real-time RT-PCR validation of microarray data for 45 genes and three housekeeping genes (HPRT1, GAPDH and B-Actin) was performed using the BioMark™ 48.48 Dynamic Arrays (Fluidigm Corporation, South San Francisco, Calif.). A sample mix, containing 2.5 μl of pre-amplified cDNA, 2.5 μl of TaqMan® Universal PCR Master Mix (Applied Biosystems, Foster City, Calif.) and 0.25 μl of DA Sample Loading Reagent (Fluidigm Corporation, South San Francisco, Calif.) and an assay mix containing 2.5 μl 20× TaqMan® Gene Expression Assay (Applied Biosystems, Foster City, Calif.) and 2.5 μl DA Assay Loading reagent (Fluidigm Corporation, South San Francisco, Calif.) were prepared. Following priming of the 48.48 Dynamic Array with control line fluid in an IFC controller (Fluidigm Corporation, South San Francisco, Calif.), 5 μl sample mix was loaded into each sample inlet and 5 μl assay mix into the detector inlet of the chip. All samples were loaded in duplicate. The chip was subsequently placed in the IFC Controller for loading and mixing of the samples and assays and then transferred to the BioMark™ Real-Time PCR System. The cycling program consisted of 10 minutes at 95° C. followed by 40 cycles of 95° C. for 15 seconds and 1 minute at 60° C.

Data was analyzed using the Fluidigm Gene Expression Data Analysis software (version 2.1.1, Fluidigm Corporation, South San Francisco, Calif.) to obtain CT values. The relative abundance was calculated according to the formula: 2^(average CT gene A−average CT HPRT1). HPRT1 was the most stable house keeping gene. Results are shown in FIGS. 10A-D.

In each of FIGS. 10A-D, box-plots for each gene in each sample are shown grouped by the molecular phenotypes, F1, F2, L, and M. Five healthy controls (Nrml) and 41 uninflamed osteoarthritis (OA) samples were included for reference. Boxes within each plot represent the 25th to 75th percentiles, horizontal lines represent medians, whiskers represent estimates of the 95% confidence intervals and individual dots correspond to each observation.

The results for the F1 phenotype are shown in FIG. 10A. Periostin (POSTN) was validated as an F1-specific transcript (FIG. 10A). Others have shown that POSTN is expressed predominantly in collagen-rich fibrous connective tissues that are subjected to constant mechanical stresses (Oku et al., Int J Hematol. 88(1):57-63 (2008)). POSTN expression is induced by TGFB and has been shown to be a component of bone marrow fibrosis. Additional validated F1-specific transcripts included ADAM12, CTHRC1, and ENPEP (FIG. 10A)

As shown in FIG. 10B, the F2-specific transcripts BTC, CLU, CRLF1 and TIMP3 were all upregulated in patient samples identified as F2, however, the levels of these transcripts in F2 tissues were not significantly different from the levels found in OA tissues.

As indicated in FIG. 10C, the B cell transcripts CD19, TNFRSF7, IgJ and IRTA2 demonstrated specificity for the L phenotype. Expression of each of these transcripts in L tissues was significantly higher than in normal and OA tissues.

As shown in FIG. 10D, CCL2, CXCL3 and VEGFA were all specific to the M phenotype, whereas the macrophage activation marker CD68 showed similar levels in all phenotypes except F1 which had similar low levels with normal controls. Also noteworthy were the levels of VEGFA, which was unique to the M phenotype among RA and OA samples, but did not differ from normal samples.

These findings provide platform-independent validation of phenotype-specific differential gene expression. Importantly, all of the analytes tested here encode for cell surface and/or soluble proteins and could therefore serve as phenotype-specific biomarkers that may be measurable systemically or in synovial fluid. In addition, because these analytes were easily detectable by qPCR, the possibility of direct synovial tissue assessment could be feasible using minimally invasive biopsy techniques.

Example 3

As described above, the L subtype was associated with the presence of organized lymphoid structures in histological sections of synovial tissue. These lymphoid clusters were also shown to contain large numbers of B cells (see, e.g., FIG. 3H). Furthermore, it is likely that the lymphoid clusters contain antibody secreting plasma cells based on the morphology of the clusters which resemble germinal centers. In addition, as described above, the L subtype was associated with the expression of genes characteristic of B cells, plasma cells, and other cells. Such genes include, as indicated in Table 1, IRTA2 (FcRH5) and CXCL13. While CXCL13 is a soluble chemokine that can be detected systemically, full length FcRH5 is a B-cell-restricted membrane bound protein. It is also, however, expressed as a truncated soluble protein due to alternate splicing of the primary mRNA (Hatzivassiliou, G., et al., Immunity 14:277-289, doi:S1074-7613(01)00109-1 [pii] (2001); Ise, T., et al., Leukemia 21:169-174, doi:2404445 [pii] 10.1038/sj.leu.2404445 (2007)). Accordingly, we hypothesized that sFcRH5 and CXCL13 might be measurable in the serum of RA patients and if so, could prove useful as serum biomarkers of the L subtype. Moreover, because a number of therapeutic agents target B cells, including anti-CD20 therapeutic antibodies, such as rituximab, we sought to determine whether serum sFcRH5 and/or CXCL13 could be useful as biomarkers for predicting patient responsiveness to such therapeutic agents.

We thus conducted the following experiments to ascertain whether serum sFcRH5 and CXCL13 levels could be used as biomarkers of the L subtype of RA and/or to predict patient responsiveness to anti-B cell therapeutic agents. As an exemplary anti-B cell therapeutic agent, we chose rituximab. Serum from 339 RA patients in a double blind, placebo-controlled phase III randomized controlled trial known as REFLEX (Randomized Evaluation of Long-Term Efficacy of Rituximab in RA) was collected and analyzed as described further below. The REFLEX trial was conducted by Genentech, Inc., Biogen-Idec, Inc. and Roche, the topline clinical findings of which were published by Cohen, S. B., et al., Arthritis Rheum 54:2793-2806 (2006).

First, we assayed levels of sFcRH5 in the patient sera at baseline (one day prior to dosing with rituximab) and compared that to levels in healthy control samples. To assay sFcRH5, we used an anti-FcRH5 monoclonal antibody, 6H1, (ATCC No. PTA-7211) that recognizes the extracellular domain of the FcRH5 molecule. This antibody is also described in International Patent Application No. PCT/US2010/029516. ELISA wells (384/plate) were coated with ms6H1 mAb at 0.5 μg/mL in 0.05M Carbonate/Bicarbonate buffer (pH 9.6) at 2-8° C. overnight. After removal of coat solution, nonspecific binding sites were blocked by incubating for at least 1 hr with blocking solution (PBS/0.5% BSA/0.05% Tween20/15 ppm Proclin, 50 μl/well). After washing the plates with 100 μl wash buffer (PBS/0.05% Tween), standard (20-0.156 ng/ml) or sample diluted in assay buffer (PBS/0.5% BSA/0.05% Tween-20/15 ppm Proclin 300/0.25% CHAPS/0.35M NaCl/5 mM EDTA, pH 7.4, 5% Fetal bovine serum) was added (25 pd/well) and incubated for 2 hrs at RT then moved to 2-8° C. for overnight incubation. After an overnight incubation, the plates were allowed to shake at room temperature (RT) for 1 hr. Then the plates were washed and 70 ng/mL of biotinylated pAb from R&D Systems was added (25 μl/well) and incubated for an additional 1 hr. Following washing, streptavidin-horseradish peroxidase (Amdex) diluted 1:10,000 was added to the plate, and incubated for 30 min. Following another wash, tetramethyl benzidine substrate (Moss TMB) was added (25 μL/well), color was allowed to develop for 15 min, and the reaction was stopped by the addition of 1 M phosphoric acid (25 μl/well). The plates were read at a wavelength of 450 nm, with reference at 630 nm, using a microplate reader (Thermo Labsystems, Finland). The concentration of soluble FcRH5 in the samples was extrapolated from a 4-parameter fit of the standard curve.

As shown in FIG. 11A, the serum level of sFcRH5 was clearly elevated in some RA patients compared to healthy controls. Accordingly, these results support the hypothesis that sFcRH5 serum levels can be used as a serum biomarker of RA, including L subtype of RA.

In addition, we determined the serum levels of CXCL13 in the same patient samples and healthy controls using the human CXCL13/BLC/BCA-1 Quantikine ELISA Kit from R&D Systems (Cat. No. DCX130). The data are shown in FIG. 11B. Similar to the results with sFcRH5, FIG. 11B shows that serum levels of CXCL13 were elevated in some RA patients compared to healthy controls. Thus, these results support the hypothesis that CXCL13 serum levels can be used as a serum biomarker of RA, including L subtype of RA.

Next, we conducted a threshold sensitivity analysis of the sFcRH5 and CXCL13 data to identify patient subgroups within the REFLEX trial with greater clinical benefit to rituximab as defined by ACR50 response at 24 weeks. The threshold sensitivity analysis was conducted as follows. The objective was to identify candidate biomarker subgroups that represented at least 20% of patients from the REFLEX trial and enriched for placebo-corrected ACR50 responses (ACR50 for the rituximab plus methotrexate group minus ACR50 for the placebo plus methotrexate group) at week 24 after the first course of rituximab. To identify subgroups with increased clinical benefit, the study population from REFLEX was stratified using baseline clinical characteristics and serological biomarkers measured in patients for whom serum samples were available. The baseline characteristics for the patient subgroups that had matching biomarker serum samples were comparable with the overall patient group in the clinical trial. For surveys of each continuous biomarker (where a range of discrete values was possible) and outcome measure ACR50 at week 24, a plot was generated presenting subgroup efficacy differentials versus a range of potential threshold values (20th-80^(th) biomarker percentiles in 5-percentile increments) to control bias. The threshold giving the largest efficacy differential (Δhigh−Δlow) was then identified. For this threshold, a permutation test was used to address statistical significance. For each permutation, biomarker values were permuted and both treatment assignment and the outcome measure were fixed. The largest efficacy differential was computed for the permutated data set, which was compared to the largest efficacy differential observed from the original data. Permutation p-values were based on 2000 permutations. A 95% confidence interval on the largest efficacy differential was calculated.

FIG. 12 shows that a subgroup of patients with sFcRH5 levels greater than 126.7 ng/ml, and a subgroup of patients with CXCL13 levels greater than 116.6 pg/ml, demonstrated significantly higher ACR50 response rates compared with patients with lower levels of these biomarkers. The striped bars in FIG. 12 are rituximab-treated patients. FIG. 12 also shows that the placebo response rates (open bars) for these biomarker-defined subgroups did not behave in a similar manner. The right side of FIG. 12 shows the optimal subgroup efficacy difference (with 95% CI); i.e., the placebo-corrected efficacy difference between the biomarker-high and biomarker-low subgroups. Because the biomarker levels of both CXCL13 and sFcRH5 were associated with improved ACR50 rates in rituximab-treated patients, but not in placebo-treated patients, these results suggest that CXCL13 and sFcRH5 serum levels are predictive of enhanced responsiveness to rituximab and not, for example, simply prognostic for disease severity and/or progression.

We next assessed the level of the putative lymphoid signature serum marker Rheumatoid Factor (RF), a prototypical RA autoantibody, in combination with sFcRH5 in serum of patients in the REFLEX trial (described above) or a second trial known as SERENE. SERENE (Study Evaluating Rituximab's Efficacy in MTX iNadequate rEsponders) was also a pivotal placebo-controlled clinical trial of rituximab, but in DMARD-IR RA patients, the topline clinical findings of which were published by Emery et al., Ann Rheum Dis. 69(9):1629-35 (2010). In these experiments, sFcRH5 was assayed as described above. RF was assayed using a commercially available ELISA kit that measures IgM, IgG, and IgA isotypes of RF (Catalog #303-305, TheraTest Labs, Lombard, Ill.).

FIG. 13 shows the results of the assessment of placebo-controlled 24 week ACR50 response rates in lymphoid biomarker-defined patient subsets in both the REFLEX (FIG. 13A) and SERENE (FIG. 13B) trials. ACR50 response rates are shown for all patients in each study as well as patient subsets defined by sFcRH5 level and/or seropositivity for RF. The concentration cutoffs for sFcRH5 high vs sFcRH5 low subsets were 126.7 ng/ml for REFLEX and 165 ng/ml for SERENE as determined by a threshold sensitivity analysis for each study. The threshold sensitivity analysis was conducted as described above. The numbers of patients treated with rituximab or placebo, as well as the numbers of patients that subsequently met the ACR50 response criteria, are shown in FIGS. 13A-B for each subset. The placebo-controlled ACR50 response rate (ΔACR50) is indicated for each subset. As can be seen in FIGS. 13A and 13B, patient subsets with elevated sFcRH5 and also seropositive for RF had enhanced placebo-controlled ACR50 response rates compared with the unselected trial population. In contrast, the patient subsets defined by low sFcRH5 levels and/or seronegative status for RF had diminished placebo-controlled ACR50 rates.

In addition, in the REFLEX study we examined at baseline the soluble FcRH5 and RF biomarkers in combination with serum levels of CXCL13 for which an optimal cut-point (116.6 pg/ml) had been determined using the threshold sensitivity method (see above). RF, sFcRH5 and CXCL13 were assayed as described above. FIG. 14 shows that patients with low baseline levels of all three biomarkers have no ACR50 response to rituximab treatment, while patients with high levels of all three biomarkers have enriched response to rituximab treatment. These data suggest that activity of the B cell pathway, a hallmark of the lymphoid subset, influences subsequent clinical response to B cell depletion therapy.

In summary, these data support the hypothesis that patients with RA characterized by a lymphoid infiltrate in their tissues, and with elevated serum levels of biomarkers specifically and significantly expressed in the L subtype gene expression signature, i.e., sFcRH5, CXCL13, and RF have a more robust clinical response to a B cell-depleting agent such as rituximab.

TABLE 5 Gene expression associated with the L subtype of RA. NETAFFX: Fold ProbeID NETAFFX: GeneTitle GeneSymbol Change p-value 221286_s_at proapoptotic caspase adaptor protein PACAP 22.72 1E−15 223565_at proapoptotic caspase adaptor protein PACAP 14.29 5E−18 204698_at interferon stimulated gene 20 kDa ISG20 9.35 2E−09 1552623_at hematopoietic SH2 domain containing HSH2D 9.32 1E−13 205267_at “POU domain, class 2, associating factor 1” POU2AF1 8.99 6E−21 228592_at “Membrane-spanning 4-domains, subfamily A, member 1” MS4A1 8.48 1E−12 206641_at “tumor necrosis factor receptor superfamily, member 17” TNFRSF17 6.79 4E−22 221602_s_at regulator of Fas-induced apoptosis | regulator of Fas-induced apoptosis TOSO 6.43 5E−11 215946_x_at “immunoglobulin lambda-like polypeptide 1 | similar to bK246H3.1 IGLL1 | 6.37 2E−18 (immunoglobulin lambda-like polypeptide 1, pre-B-cell specific)” LOC91316 204269_at pim-2 oncogene PIM2 6.27 1E−18 235401_s_at Fc receptor homolog expressed in B cells FREB 6.21 2E−14 230673_at polycystic kidney and hepatic disease 1 (autosomal recessive)-like 1 PKHD1L1 6.16 2E−12 219463_at chromosome 20 open reading frame 103 C20orf103 6.14 3E−11 219159_s_at SLAM family member 7 SLAMF7 6.07 2E−18 217231_s_at microtubule associated serine/threonine kinase 1 MAST1 5.85 3E−12 217418_x_at “membrane-spanning 4-domains, subfamily A, member 1” MS4A1 5.71 7E−13 229971_at G protein-coupled receptor 114 GPR114 5.38 4E−19 230075_at “RAB39B, member RAS oncogene family” RAB39B 5.34 9E−15 1552999_a_at WAP four-disulfide core domain 10B WFDC10B 5.28 7E−12 210356_x_at “membrane-spanning 4-domains, subfamily A, member 1” MS4A1 5.18 1E−13 205544_s_at complement component (3d/Epstein Barr virus) receptor 2 CR2 5.12 3E−09 222838_at SLAM family member 7 SLAMF7 4.99 3E−20 234306_s_at SLAM family member 7 SLAMF7 4.80 9E−15 238695_s_at “RAB39B, member RAS oncogene family” RAB39B 4.78 3E−13 235400_at Fc receptor homolog expressed in B cells FREB 4.65 4E−12 228599_at “Membrane-spanning 4-domains, subfamily A, member 1” MS4A1 4.63 4E−13 210370_s_at lymphocyte antigen 9 LY9 4.51 1E−15 206866_at “cadherin 4, type 1, R-cadherin (retinal)” CDH4 4.50 4E−09 232112_at Ral GEF with PH domain and SH3 binding motif 2 RALGPS2 4.43 4E−14 220338_at Ral GEF with PH domain and SH3 binding motif 2 RALGPS2 4.40 7E−17 237759_at CD48 antigen (B-cell membrane protein) CD48 4.30 2E−13 241844_x_at hypothetical protein FLJ23235 FLJ23235 4.29 7E−15 201689_s_at tumor protein D52 TPD52 4.27 2E−16 236280_at Transcribed locus NA 4.26 2E−17 226147_s_at polymeric immunoglobulin receptor PIGR 4.21 2E−06 1553039_a_at ankyrin repeat and SOCS box-containing 10 ASB10 4.19 2E−12 201688_s_at tumor protein D52 TPD52 4.16 9E−15 206296_x_at mitogen-activated protein kinase kinase kinase kinase 1 MAP4K1 4.12 4E−08 212311_at KIAA0746 protein KIAA0746 4.10 8E−22 229152_at chromosome 4 open reading frame 7 C4orf7 4.08 6E−06 1555613_a_at zeta-chain (TCR) associated protein kinase 70 kDa ZAP70 4.06 1E−12 215967_s_at lymphocyte antigen 9 LY9 4.06 8E−11 215779_s_at “histone 1, H2bg” HIST1H2BG 4.06 6E−11 219888_at sperm associated antigen 4 SPAG4 3.90 6E−17 204960_at “protein tyrosine phosphatase, receptor type, C-associated protein” PTPRCAP 3.87 2E−14 205753_at “C-reactive protein, pentraxin-related” CRP 3.86 1E−16 223750_s_at toll-like receptor 10 TLR10 3.86 2E−08 211517_s_at “interleukin 5 receptor, alpha” IL5RA 3.82 5E−10 235863_at homolog of mouse skeletal muscle sarcoplasmic reticulum FLJ32416 3.75 4E−09 protein JP-45 214469_at “histone 1, H2ae” HIST1H2AE 3.64 2E−08 227189_at copine V CPNE5 3.62 3E−21 229721_x_at “Der1-like domain family, member 3” DERL3 3.61 1E−19 201691_s_at tumor protein D52 TPD52 3.56 8E−14 220059_at BCR downstream signaling 1 BRDG1 3.54 3E−14 211789_s_at Mlx interactor MONDOA 3.50 4E−08 201690_s_at tumor protein D52 TPD52 3.49 5E−18 210279_at G protein-coupled receptor 18 GPR18 3.49 2E−12 206686_at “pyruvate dehydrogenase kinase, isoenzyme 1” PDK1 3.42 5E−13 207245_at “UDP glycosyltransferase 2 family, polypeptide B17” UGT2B17 3.41 4E−06 1557719_at “phosphatidylinositol-3-phosphate/phosphatidylinositol 5-kinase, type III” PIP5K3 3.35 1E−22 220035_at nucleoporin 210 kDa NUP210 3.34 4E−13 221601_s_at regulator of Fas-induced apoptosis | regulator of Fas-induced apoptosis TOSO 3.32 7E−11 214615_at “purinergic receptor P2Y, G-protein coupled, 10” P2RY10 3.27 1E−14 225792_at Hook homolog 1 (Drosophila) HOOK1 3.24 4E−16 234383_x_at NA NA 3.23 2E−12 205903_s_at “potassium intermediate/small conductance calcium-activated KCNN3 3.22 2E−08 channel, subfamily N, member 3” 240070_at Hypothetical protein FLJ39873 FLJ39873 3.21 9E−13 1555342_a_at unc-5 homolog C (C. elegans) UNC5C 3.20 8E−15 206121_at adenosine monophosphate deaminase 1 (isoform M) AMPD1 3.20 1E−12 229629_at Transcribed locus NA 3.17 4E−12 223246_s_at spermatid perinuclear RNA binding protein STRBP 3.15 2E−11 242458_at Ral GEF with PH domain and SH3 binding motif 2 RALGPS2 3.14 5E−13 240098_at RAP1 interacting factor homolog (yeast) RIF1 3.12 5E−10 210448_s_at “purinergic receptor P2X, ligand-gated ion channel, 5” P2RX5 3.06 9E−12 204891_s_at lymphocyte-specific protein tyrosine kinase LCK 3.03 2E−09 240265_at TRAF3-interacting Jun N-terminal kinase (JNK)-activating modulator T3JAM 3.01 9E−10 228705_at calpain 12 CAPN12 2.99 2E−09 215243_s_at “gap junction protein, beta 3, 31 kDa (connexin 31)” GJB3 2.99 1E−13 207237_at “potassium voltage-gated channel, shaker-related subfamily, member 3” KCNA3 2.98 8E−11 228897_at “Der1-like domain family, member 3” DERL3 2.95 2E−16 213888_s_at TRAF3-interacting Jun N-terminal kinase (JNK)-activating modulator T3JAM 2.90 1E−13 243969_at “solute carrier family 24 (sodium/potassium/calcium exchanger), SLC24A4 2.90 3E−10 member 4” 209602_s_at GATA binding protein 3 GATA3 2.89 9E−08 206398_s_at CD19 antigen CD19 2.87 3E−16 220169_at hypothetical protein FLJ23235 FLJ23235 2.86 9E−14 237251_at hypothetical protein FLJ32884 FLJ32884 2.85 2E−08 223751_x_at toll-like receptor 10 TLR10 2.83 1E−14 235165_at par-6 partitioning defective 6 homolog beta (C. elegans) PARD6B 2.83 8E−08 227134_at synaptotagmin-like 1 SYTL1 2.82 2E−20 208553_at “histone 1, H1e” HIST1H1E 2.82 9E−05 206780_at “glutamate decarboxylase 2 (pancreatic islets and brain, 65 kDa)” GAD2 2.82 2E−05 206048_at zinc finger protein 339 ZNF339 2.82 1E−08 200670_at X-box binding protein 1 XBP1 2.81 7E−22 220507_s_at “ureidopropionase, beta” UPB1 2.81 5E−13 231124_x_at Lymphocyte antigen 9 LY9 2.81 6E−13 209829_at chromosome 6 open reading frame 32 C6orf32 2.80 1E−08 224102_at “purinergic receptor P2Y, G-protein coupled, 12” P2RY12 2.80 2E−05 222067_x_at “histone 1, H2bd” HIST1H2BD 2.78 8E−15 233500_x_at “C-type lectin superfamily 2, member D” CLEC2D 2.77 5E−08 1561820_at “sodium channel, voltage gated, type VIII, alpha” SCN8A 2.76 6E−15 241914_s_at hypothetical protein LOC123876 | xenobiotic/medium-chain fatty LOC123876 | 2.76 5E−13 acid: CoA ligase HXMA 219014_at placenta-specific 8 PLAC8 2.76 2E−11 228167_at kelch-like 6 (Drosophila) KLHL6 2.75 2E−19 206896_s_at “guanine nucleotide binding protein (G protein), gamma 7” GNG7 2.74 4E−16 216277_at BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) BUB1 2.74 1E−12 228258_at FLJ00332 protein FLJ00332 2.72 2E−11 1553237_x_at “protocadherin alpha subfamily C, 1” PCDHAC1 2.72 4E−10 214470_at “killer cell lectin-like receptor subfamily B, member 1 | killer cell KLRB1 2.71 4E−12 lectin-like receptor subfamily B, member 1” 220126_at testes-specific protease 50 TSP50 2.71 1E−07 204563_at selectin L (lymphocyte adhesion molecule 1) SELL 2.70 1E−09 1560219_at “CDNA FLJ34490 fis, clone HLUNG2004707” NA 2.67 4E−13 210538_s_at baculoviral IAP repeat-containing 3 BIRC3 2.67 1E−10 237323_at hypothetical protein FLJ22761 FLJ22761 2.66 2E−10 204949_at intercellular adhesion molecule 3 ICAM3 2.65 3E−17 212314_at KIAA0746 protein KIAA0746 2.65 3E−18 224520_s_at hypothetical protein MGC13168 | hypothetical protein MGC13168 MGC13168 2.65 1E−10 228320_x_at hypothetical protein LOC92558 LOC92558 2.65 1E−09 206255_at B lymphoid tyrosine kinase BLK 2.64 2E−08 1558972_s_at chromosome 6 open reading frame 190 C6orf190 2.64 2E−09 1555086_at signal transducer and activator of transcription 5B STAT5B 2.63 9E−08 237159_x_at “Transcribed locus, weakly similar to XP_517655.1 similar to NA 2.62 2E−08 KIAA0825 protein [Pan troglodytes]” 206337_at chemokine (C-C motif) receptor 7 | chemokine (C-C motif) receptor 7 CCR7 2.62 5E−11 1552596_at growth arrest-specific 2 like 2 GAS2L2 2.61 2E−12 230011_at similar to mouse meiosis defective 1 gene MGC40042 2.60 9E−17 226452_at “pyruvate dehydrogenase kinase, isoenzyme 1” PDK1 2.59 7E−16 229686_at “purinergic receptor P2Y, G-protein coupled, 8” P2RY8 2.59 6E−19 214540_at “histone 1, H2bo” HIST1H2BO 2.58 4E−09 205890_s_at ubiquitin D UBD 2.58 2E−07 223299_at SEC11-like 3 (S. cerevisiae) SEC11L3 2.58 3E−16 239088_at Transcribed locus NA 2.58 2E−10 1552940_at putative membrane protein HE9 HE9 2.58 2E−06 230863_at Low density lipoprotein-related protein 2 LRP2 2.58 2E−06 1563290_at “dynein, axonemal, heavy polypeptide 3” DNAH3 2.57 7E−11 213915_at natural killer cell group 7 sequence NKG7 2.55 7E−09 208515_at “histone 1, H2bm” HIST1H2BM 2.55 3E−10 209813_x_at T cell receptor gamma variable 9 | T cell receptor gamma TRGV9 | 2.55 2E−12 variable 9 | TCR gamma alternate reading frame protein | TARP TCR gamma alternate reading frame protein 219551_at ELL associated factor 2 EAF2 2.54 2E−18 216901_s_at “zinc finger protein, subfamily 1A, 1 (Ikaros)” ZNFN1A1 2.54 3E−07 1554069_at EPH receptor A8 EPHA8 2.53 7E−07 237230_at glycoprotein hormone alpha 2 GPHA2 2.53 1E−05 1558971_at chromosome 6 open reading frame 190 C6orf190 2.52 3E−12 232352_at “ISL2 transcription factor, LIM/homeodomain, (islet-2)” ISL2 2.52 9E−07 214032_at zeta-chain (TCR) associated protein kinase 70 kDa ZAP70 2.51 2E−09 1565836_at NA NA 2.48 1E−10 231746_at Mix1 homeobox-like 1 (Xenopus laevis) MIXL1 2.48 2E−11 241860_at Serine/threonine kinase 17b (apoptosis-inducing) STK17B 2.48 4E−11 221271_at interleukin 21 IL21 2.47 2E−07 244780_at sphingosine-1-phosphate phosphotase 2 SGPP2 2.46 1E−06 231021_at “Solute carrier family 6 (neurotransmitter transporter), member 19” SLC6A19 2.46 2E−08 1554240_a_at “integrin, alpha L (antigen CD11A (p180), lymphocyte ITGAL 2.45 4E−12 function-associated antigen 1; alpha polypeptide)” 1555687_a_at “C-type lectin domain family 4, member C” CLEC4C 2.44 4E−09 217073_x_at apolipoprotein A-I APOA1 2.43 7E−05 243797_at serine/threonine kinase 17b (apoptosis-inducing) STK17B 2.42 2E−11 215960_at “solute carrier family 5 (low affinity glucose cotransporter), SLC5A4 2.42 2E−07 member 4” 204536_s_at NA NA 2.42 2E−07 1553279_at butyrophilin-like 9 BTNL9 2.41 2E−05 1553856_s_at “purinergic receptor P2Y, G-protein coupled, 10” P2RY10 2.40 6E−14 236341_at cytotoxic T-lymphocyte-associated protein 4 CTLA4 2.40 7E−09 211181_x_at runt-related transcription factor 1 (acute myeloid leukemia 1; RUNX1 2.39 3E−08 aml1 oncogene) 213534_s_at PAS domain containing serine/threonine kinase PASK 2.39 2E−11 208546_x_at “histone 1, H2bh” HIST1H2BH 2.38 6E−07 222897_s_at zinc finger protein 64 homolog (mouse) ZFP64 2.38 3E−09 235372_at Fc receptor homolog expressed in B cells FREB 2.37 9E−16 224538_s_at par-6 partitioning defective 6 homolog gamma (C. elegans) | PARD6G 2.37 6E−12 par-6 partitioning defective 6 homolog gamma (C. elegans) 205804_s_at TRAF3-interacting Jun N-terminal kinase (JNK)-activating modulator T3JAM 2.37 1E−09 229360_at suppressor of hairy wing homolog 2 (Drosophila) SUHW2 2.36 3E−11 1554343_a_at BCR downstream signaling 1 BRDG1 2.36 1E−10 1552386_at hypothetical protein FLJ33641 FLJ33641 2.36 6E−10 237162_at ankyrin repeat domain 15 ANKRD15 2.35 1E−05 1552839_at chromosome 14 open reading frame 54 C14orf54 2.35 5E−07 221331_x_at cytotoxic T-lymphocyte-associated protein 4 CTLA4 2.34 8E−11 233252_s_at spermatid perinuclear RNA binding protein STRBP 2.34 3E−18 210769_at cyclic nucleotide gated channel beta 1 CNGB1 2.33 2E−07 204470_at “chemokine (C-X-C motif) ligand 1 (melanoma growth CXCL1 2.33 2E−06 stimulating activity, alpha)” 204475_at matrix metalloproteinase 1 (interstitial collagenase) MMP1 2.33 8E−06 205884_at “integrin, alpha 4 (antigen CD49D, alpha 4 subunit of VLA-4 receptor)” ITGA4 2.31 9E−11 223953_s_at zinc finger and BTB domain containing 37 ZBTB37 2.30 3E−05 207635_s_at “potassium voltage-gated channel, subfamily H (eag-related), member 1” KCNH1 2.30 1E−06 1553890_s_at hypothetical protein BC018697 LOC126147 2.30 6E−12 211469_s_at chemokine (C-X-C motif) receptor 6 CXCR6 2.29 6E−09 220286_at hypothetical protein FLJ20313 FLJ20313 2.29 2E−05 204890_s_at lymphocyte-specific protein tyrosine kinase LCK 2.29 2E−09 228298_at hypothetical protein MGC16044 MGC16044 2.29 2E−17 216920_s_at T cell receptor gamma variable 9 | TCR gamma alternate reading TRGV9 | 2.29 1E−16 frame protein TARP 214595_at “potassium voltage-gated channel, subfamily G, member 1” KCNG1 2.28 5E−07 228236_at chromosome 20 open reading frame 54 C20orf54 2.28 3E−07 221690_s_at “NACHT, leucine rich repeat and PYD containing 2” NALP2 2.28 1E−12 208523_x_at “histone 1, H2bi” HIST1H2BI 2.27 1E−05 221442_at melanocortin 3 receptor MC3R 2.26 4E−13 233504_at chromosome 9 open reading frame 84 C9orf84 2.26 4E−05 1553675_at kinesin-like 8 KNSL8 2.26 3E−07 205309_at “sphingomyelin phosphodiesterase, acid-like 3B” SMPDL3B 2.25 1E−10 211339_s_at IL2-inducible T-cell kinase ITK 2.25 1E−08 206367_at renin REN 2.25 2E−07 244033_at NA NA 2.25 3E−18 232635_at chromosome 14 open reading frame 145 C14orf145 2.24 1E−14 227353_at Epidermodysplasia verruciformis 2 EVER2 2.24 3E−12 1552806_a_at sialic acid binding Ig-like lectin 10 SIGLEC10 2.23 1E−06 33304_at interferon stimulated gene 20 kDa ISG20 2.23 7E−14 206369_s_at “phosphoinositide-3-kinase, catalytic, gamma polypeptide” PIK3CG 2.22 3E−06 238629_x_at NA NA 2.22 4E−06 205254_x_at “transcription factor 7 (T-cell specific, HMG-box)” TCF7 2.22 4E−10 1553804_a_at hypothetical protein FLJ25414 FLJ25414 2.21 4E−11 204118_at CD48 antigen (B-cell membrane protein) | CD48 antigen CD48 2.20 3E−15 (B-cell membrane protein) 211885_x_at “fucosyltransferase 6 (alpha (1,3) fucosyltransferase)” FUT6 2.20 3E−07 1569980_x_at GLI-Kruppel family member HKR1 HKR1 2.20 3E−05 1552682_a_at AF15q14 protein AF15Q14 2.20 2E−07 215806_x_at T cell receptor gamma variable 9 | TCR gamma alternate TRGV9 | 2.20 8E−15 reading frame protein TARP 223903_at toll-like receptor 9 TLR9 2.20 5E−16 206785_s_at “killer cell lectin-like receptor subfamily C, member 1 | KLRC1 | 2.20 4E−11 killer cell lectin-like receptor subfamily C, member 2” KLRC2 217552_x_at “complement component (3b/4b) receptor 1, including Knops CR1 2.20 1E−07 blood group system” 216945_x_at PAS domain containing serine/threonine kinase PASK 2.19 2E−10 211182_x_at runt-related transcription factor 1 (acute myeloid leukemia 1; RUNX1 2.19 2E−07 aml1 oncogene) 1556149_at armadillo repeat gene deletes in velocardiofacial syndrome ARVCF 2.19 9E−13 210265_x_at NA NA 2.19 1E−05 227182_at sushi domain containing 3 SUSD3 2.18 1E−09 228795_at “Protein kinase C, beta 1” PRKCB1 2.18 4E−08 1570200_at helicase (DNA) B HELB 2.17 8E−07 206324_s_at death-associated protein kinase 2 DAPK2 2.17 3E−09 236554_x_at epidermodysplasia verruciformis 2 EVER2 2.16 3E−13 233094_at “Ubiquitin-conjugating enzyme E2D 1 (UBC4/5 homolog, yeast)” UBE2D1 2.16 5E−07 228865_at specifically androgen-regulated protein SARG 2.16 7E−08 225619_at hypothetical protein FLJ30046 FLJ30046 2.15 9E−10 210031_at “CD3Z antigen, zeta polypeptide (TiT3 complex)” CD3Z 2.15 2E−11 205885_s_at “integrin, alpha 4 (antigen CD49D, alpha 4 subunit of VLA-4 receptor)” ITGA4 2.15 1E−09 219852_s_at hypothetical protein FLJ13941 FLJ13941 2.15 7E−07 209398_at “histone 1, H1c” HIST1H1C 2.15 1E−11 209353_s_at hypothetical protein MGC16664 MGC16664 2.14 4E−06 228360_at hypothetical protein LOC130576 LOC130576 2.14 8E−09 215925_s_at CD72 antigen CD72 2.14 2E−09 207849_at interleukin 2 IL2 2.14 6E−05 213475_s_at “integrin, alpha L (antigen CD11A (p180), lymphocyte ITGAL 2.14 6E−10 function-associated antigen 1; alpha polypeptide)” 227641_at F-box and leucine-rich repeat protein 16 FBXL16 2.14 6E−15 236995_x_at Transcription factor EC TFEC 2.13 4E−10 1553057_at “serine (or cysteine) proteinase inhibitor, clade B (ovalbumin), SERPINB12 2.13 1E−07 member 12” 208574_at SRY (sex determining region Y)-box 14 SOX14 2.13 2E−06 214567_s_at chemokine (C motif) ligand 1 | chemokine (C motif) ligand 2 XCL1 | XCL2 2.12 7E−06 238531_x_at NA NA 2.12 5E−08 231966_at “protein phosphatase 1, regulatory (inhibitor) subunit 9A” PPP1R9A 2.12 1E−05 244578_at lymphocyte cytosolic protein 2 (SH2 domain containing leukocyte LCP2 2.12 6E−06 protein of 76 kDa) 244781_x_at IBR domain containing 2 IBRDC2 2.12 6E−06 223709_s_at “wingless-type MMTV integration site family, member 10A” WNT10A 2.12 7E−14 236655_at Tumor protein D52 TPD52 2.11 4E−11 211232_x_at glucagon-like peptide 1 receptor GLP1R 2.11 2E−08 207226_at “histone 1, H2bn” HIST1H2BN 2.11 7E−05 204852_s_at “protein tyrosine phosphatase, non-receptor type 7” PTPN7 2.11 7E−10 206437_at “endothelial differentiation, G-protein-coupled receptor 6” EDG6 2.11 3E−09 1552552_s_at “C-type lectin domain family 4, member C” CLEC4C 2.10 2E−11 1560225_at “Homo sapiens, clone IMAGE: 5244076, mRNA” NA 2.10 9E−08 231776_at eomesodermin homolog (Xenopus laevis) EOMES 2.09 2E−08 212316_at nucleoporin 210 kDa NUP210 2.09 8E−15 206244_at “complement component (3b/4b) receptor 1, including CR1 2.09 8E−08 Knops blood group system” 209695_at “protein tyrosine phosphatase type IVA, member 3” PTP4A3 2.09 2E−08 224273_at chromosome 3 open reading frame 20 C3orf20 2.09 1E−11 201287_s_at syndecan 1 SDC1 2.08 2E−09 240254_at TRAF2 and NCK interacting kinase TNIK 2.08 2E−08 209603_at GATA binding protein 3 GATA3 2.08 6E−05 222859_s_at dual adaptor of phosphotyrosine and 3-phosphoinositides DAPP1 2.07 6E−12 1552807_a_at sialic acid binding Ig-like lectin 10 SIGLEC10 2.06 1E−05 1553132_a_at membrane targeting (tandem) C2 domain containing 1 MTAC2D1 2.06 3E−08 208060_at paired box gene 7 PAX7 2.06 7E−07 223732_at “solute carrier family 23 (nucleobase transporters), member 1” SLC23A1 2.06 2E−06 235229_at “PREDICTED: Homo sapiens similar to Olfactory receptor 2I2 NA 2.05 4E−05 (LOC442197), mRNA” 210313_at “leukocyte immunoglobulin-like receptor, subfamily A ILT7 2.05 2E−06 (without TM domain), member 4” 206761_at CD96 antigen CD96 2.05 3E−13 241871_at calcium/calmodulin-dependent protein kinase IV CAMK4 2.05 5E−08 214157_at GNAS complex locus GNAS 2.05 4E−17 41577_at “protein phosphatase 1, regulatory (inhibitor) subunit 16B” PPP1R16B 2.05 3E−09 236852_at NA NA 2.03 7E−06 207651_at G protein-coupled receptor 171 GPR171 2.03 4E−06 212315_s_at nucleoporin 210 kDa NUP210 2.03 1E−12 219812_at Stromal antigen 3 STAG3 2.03 3E−12 207655_s_at B-cell linker BLNK 2.02 2E−17 206574_s_at “protein tyrosine phosphatase type IVA, member 3” PTP4A3 2.02 1E−08 206534_at “glutamate receptor, ionotropic, N-methyl D-aspartate 2A” GRIN2A 2.02 4E−07 211384_s_at “calcium-sensing receptor (hypocalciuric hypercalcemia 1, CASR 2.02 1E−06 severe neonatal hyperparathyroidism)” 226659_at differentially expressed in FDCP 6 homolog (mouse) DEF6 2.02 2E−13 224289_s_at FKSG83 FKSG83 2.01 5E−05 208401_s_at glucagon-like peptide 1 receptor GLP1R 2.01 5E−05 208888_s_at nuclear receptor co-repressor 2 NCOR2 2.01 3E−05 206897_at “P antigen family, member 1 (prostate associated)” PAGE1 2.01 2E−08 206978_at chemokine (C-C motif) receptor 2 | chemokine (C-C motif) receptor 2 CCR2 2.01 3E−10 205828_at “matrix metalloproteinase 3 (stromelysin 1, progelatinase)” MMP3 2.00 2E−06 206449_s_at mannan-binding lectin serine protease 1 (C4/C2 activating MASP1 2.00 7E−10 component of Ra-reactive factor) 217489_s_at interleukin 6 receptor IL6R 1.99 2E−09 215509_s_at BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) BUB1 1.99 7E−06 220279_at tripartite motif-containing 17 TRIM17 1.99 4E−05 207902_at “interleukin 5 receptor, alpha” IL5RA 1.99 2E−09 242375_x_at NA NA 1.98 7E−06 1561336_at deoxyribonuclease I-like 3 DNASE1L3 1.98 2E−07 1564713_a_at forkhead box N4 FOXN4 1.98 3E−15 235522_at NA NA 1.98 9E−12 202064_s_at sel-1 suppressor of lin-12-like (C. elegans) SEL1L 1.98 5E−10 1560397_s_at kelch-like 6 (Drosophila) KLHL6 1.97 8E−07 203399_x_at pregnancy specific beta-1-glycoprotein 3 PSG3 1.97 8E−05 234362_s_at cytotoxic T-lymphocyte-associated protein 4 CTLA4 1.97 3E−12 236454_at NA NA 1.97 1E−06 214446_at “elongation factor, RNA polymerase II, 2” ELL2 1.97 4E−05 207634_at programmed cell death 1 PDCD1 1.97 2E−07 206828_at TXK tyrosine kinase TXK 1.97 3E−07 1559051_s_at chromosome 6 open reading frame 150 C6orf150 1.96 5E−11 223536_at pleckstrin and Sec7 domain containing 2 PSD2 1.96 3E−10 229526_at aquaporin 11 AQP11 1.96 1E−05 206366_x_at chemokine (C motif) ligand 2 XCL2 1.96 7E−05 229065_at “solute carrier family 35, member F3” SLC35F3 1.95 3E−07 1553486_a_at hypothetical protein FLJ39647 FLJ39647 1.95 8E−05 204769_s_at “transporter 2, ATP-binding cassette, sub-family B (MDR/TAP)” TAP2 1.95 3E−06 211670_x_at “synovial sarcoma, X breakpoint 3 | synovial sarcoma, X breakpoint 3” SSX3 1.95 2E−05 1553176_at SH2 domain-containing molecule EAT2 EAT2 1.95 4E−08 206800_at “5,10-methylenetetrahydrofolate reductase (NADPH)” MTHFR 1.95 2E−11 220563_s_at SH3 and multiple ankyrin repeat domains 1 SHANK1 1.94 2E−06 244195_at Tubby like protein 4 TULP4 1.94 8E−05 1556653_at hypothetical protein FLJ25415 FLJ25415 1.94 9E−07 228427_at F-box protein 16 FBXO16 1.94 3E−09 235116_at TNF receptor-associated factor 1 TRAF1 1.93 1E−10 226226_at transmembrane protein 45B TMEM45B 1.93 6E−06 1559474_at aortic preferentially expressed protein 1 APEG1 1.93 3E−09 1559438_at chromosome 21 open reading frame 58 C21orf58 1.93 8E−06 1568822_at GTP binding protein 5 (putative) GTPBP5 1.93 7E−06 235117_at similar to RIKEN cDNA 2510006C20 gene LOC494143 1.92 3E−13 208490_x_at “histone 1, H2bf” HIST1H2BF 1.92 1E−12 1552787_at helicase (DNA) B HELB 1.92 1E−11 214481_at “histone 1, H2am” HIST1H2AM 1.92 9E−05 1554050_at “sphingomyelin phosphodiesterase, acid-like 3B” SMPDL3B 1.92 3E−15 240997_at Hypothetical protein LOC131873 LOC131873 1.92 1E−07 220132_s_at “C-type lectin superfamily 2, member D” CLEC2D 1.92 4E−09 241723_at IQ motif containing GTPase activating protein 2 IQGAP2 1.92 8E−05 215551_at estrogen receptor 1 ESR1 1.92 8E−06 215071_s_at NA NA 1.92 2E−08 213820_s_at START domain containing 5 STARD5 1.92 1E−05 218614_at hypothetical protein FLJ10652 FLJ10652 1.91 1E−13 219144_at hypothetical protein MGC1136 MGC1136 1.91 1E−05 220030_at serine/threonine/tyrosine kinase 1 STYK1 1.91 4E−06 243981_at serine/threonine kinase 4 STK4 1.91 8E−10 1557720_s_at myosin heavy chain Myr 8 MYR8 1.91 4E−07 229391_s_at similar to RIKEN cDNA A630077B13 gene; RIKEN cDNA 2810048G17 LOC441168 1.91 2E−05 227346_at “Zinc finger protein, subfamily 1A, 1 (Ikaros)” ZNFN1A1 1.90 6E−11 1553641_a_at “testis specific, 13” TSGA13 1.90 1E−05 226150_at HTPAP protein HTPAP 1.90 3E−15 220144_s_at ankyrin repeat domain 5 ANKRD5 1.90 1E−06 207770_x_at chorionic somatomammotropin hormone 2 CSH2 1.90 1E−05 202063_s_at sel-1 suppressor of lin-12-like (C. elegans) SEL1L 1.90 5E−15 209083_at “coronin, actin binding protein, 1A” CORO1A 1.89 1E−07 221023_s_at “potassium voltage-gated channel, subfamily H (eag-related), member 6 | KCNH6 1.89 2E−08 potassium voltage-gated channel, subfamily H (eag-related), member 6” 234037_s_at Hypothetical protein FLJ11996 FLJ11996 1.89 2E−06 214617_at perforin 1 (pore forming protein) | perforin 1 (pore forming protein) PRF1 1.89 7E−11 231338_at nuclear protein in testis NUT 1.89 7E−06 222782_s_at GEM interacting protein GMIP 1.89 5E−08 205038_at “Zinc finger protein, subfamily 1A, 1 (Ikaros)” ZNFN1A1 1.89 8E−09 216610_at “solute carrier family 6 (neurotransmitter transporter, noradrenalin), SLC6A2 1.89 7E−05 member 2” 220428_at “CD207 antigen, langerin” CD207 1.88 7E−11 1559607_s_at “Guanylate binding protein family, member 6” GBP6 1.88 1E−07 217594_at “zinc finger, CCHC domain containing 11” ZCCHC11 1.88 1E−11 1553208_s_at ADP-ribosylation factor-like 10A ARL10A 1.88 4E−05 224551_s_at “spectrin, beta, non-erythrocytic 4 | spectrin, beta, non-erythrocytic 4” SPTBN4 1.88 1E−05 202062_s_at sel-1 suppressor of lin-12-like (C. elegans) SEL1L 1.87 3E−14 212699_at secretory carrier membrane protein 5 SCAMP5 1.87 5E−19 208364_at “inositol polyphosphate-4-phosphatase, type I, 107 kDa” INPP4A 1.87 3E−09 215923_s_at pleckstrin and Sec7 domain containing 4 PSD4 1.87 3E−06 242601_at hypothetical protein LOC253012 LOC253012 1.87 4E−05 230050_at BTB (POZ) domain containing 14A BTBD14A 1.87 2E−08 223634_at “RASD family, member 2” RASD2 1.87 6E−09 215085_x_at deleted in lung and esophageal cancer 1 DLEC1 1.87 1E−15 242242_at ubiquitin specific protease 6 (Tre-2 oncogene) USP6 1.87 4E−14 244029_at Copine V CPNE5 1.87 3E−14 238567_at sphingosine-1-phosphate phosphotase 2 SGPP2 1.86 4E−06 208273_at NA NA 1.86 4E−06 208604_s_at homeo box A3 | homeo box A3 HOXA3 1.86 1E−09 219976_at hook homolog 1 (Drosophila) HOOK1 1.86 1E−05 203397_s_at UDP-N-acetyl-alpha-D-galactosamine: polypeptide N- GALNT3 1.86 6E−08 acetylgalactosaminyltransferase 3 (GalNAc-T3) 1564237_at “CDNA FLJ23858 fis, clone LNG07565” NA 1.86 3E−05 210140_at cystatin F (leukocystatin) CST7 1.86 2E−06 1555834_at Ubiquitin carboxyl-terminal esterase L1 (ubiquitin thiolesterase) UCHL1 1.86 1E−05 221377_s_at recombining binding protein suppressor of hairless (Drosophila)-like RBPSUHL 1.85 1E−07 208536_s_at BCL2-like 11 (apoptosis facilitator) BCL2L11 1.85 9E−06 1552701_a_at CARD only protein COPI 1.85 1E−10 1553387_at “ataxia telangiectasia mutated (includes complementation ATM 1.85 8E−10 groups A, C and D)” 208488_s_at “complement component (3b/4b) receptor 1, including CR1 1.85 3E−09 Knops blood group system” 220883_at NA NA 1.85 7E−07 1555275_a_at kelch-like 6 (Drosophila) KLHL6 1.84 9E−12 207681_at chemokine (C-X-C motif) receptor 3 CXCR3 1.84 3E−11 1553857_at hypothetical protein FLJ37794 FLJ37794 1.84 5E−07 207238_s_at “protein tyrosine phosphatase, receptor type, C” PTPRC 1.84 1E−07 1554601_at T-cell lymphoma breakpoint associated target 1 TCBA1 1.84 2E−05 1556849_at Ring finger protein 38 RNF38 1.84 3E−07 213416_at NA NA 1.84 6E−10 1552497_a_at SLAM family member 6 SLAMF6 1.84 1E−14 1555120_at CD96 antigen CD96 1.84 4E−09 236838_at hypothetical gene supported by BC019717 LOC440360 1.84 3E−07 236337_at hypothetical protein LOC221711 LOC221711 1.84 2E−06 220384_at thioredoxin domain containing 3 (spermatozoa) TXNDC3 1.84 3E−08 201004_at “signal sequence receptor, delta (translocon-associated protein delta)” SSR4 1.84 8E−15 239588_s_at Hypothetical protein FLJ20315 FLJ20315 1.83 2E−08 211532_x_at “killer cell immunoglobulin-like receptor, two domains, short KIR2DS2 1.83 4E−05 cytoplasmic tail, 2” 210708_x_at “caspase 10, apoptosis-related cysteine protease” CASP10 1.83 1E−08 241328_at “zinc finger, matrin type 1” ZMAT1 1.83 1E−05 228377_at kelch-like 14 (Drosophila) KLHL14 1.82 1E−08 213920_at cut-like 2 (Drosophila) CUTL2 1.81 4E−06 214907_at carcinoembryonic antigen-related cell adhesion molecule R29124_1 1.81 1E−12 1555981_at hypothetical protein DKFZp762C2414 DKFZp762C2414 1.81 4E−19 1555662_s_at D-amino acid oxidase activator DAOA 1.81 3E−09 241927_x_at NA NA 1.81 4E−08 1558698_at zinc finger protein 264 ZNF264 1.81 4E−06 203331_s_at NA NA 1.81 4E−07 38149_at NA NA 1.81 1E−15 221441_at goosecoid-like GSCL 1.81 2E−05 215586_at “Protein phosphatase 3 (formerly 2B), catalytic subunit, PPP3CB 1.81 1E−07 beta isoform (calcineurin A beta)” 226099_at “elongation factor, RNA polymerase II, 2” ELL2 1.81 1E−08 206566_at “solute carrier family 7 (cationic amino acid transporter, SLC7A1 1.80 2E−08 y+ system), member 1” 236134_at WD-repeat protein HAN11 1.80 1E−07 1556655_s_at “CDNA FLJ38740 fis, clone KIDNE2011782” NA 1.80 2E−05 242752_at MRNA (clone ICRFp507I1077) NA 1.80 4E−12 211620_x_at runt-related transcription factor 1 (acute myeloid leukemia 1; RUNX1 1.80 5E−10 aml1 oncogene) | runt-related transcription factor 1 (acute myeloid leukemia 1; aml1 oncogene) 244313_at Transcribed locus NA 1.80 8E−06 212750_at “protein phosphatase 1, regulatory (inhibitor) subunit 16B” PPP1R16B 1.80 7E−08 224285_at G protein-coupled receptor 174 GPR174 1.80 2E−07 1554834_a_at Ras association (RaIGDS/AF-6) domain family 5 RASSF5 1.79 1E−07 231549_at hypothetical protein MGC35194 MGC35194 1.79 2E−05 210712_at lactate dehydrogenase A-like 6B LDHAL6B 1.79 1E−06 219290_x_at dual adaptor of phosphotyrosine and 3-phosphoinositides DAPP1 1.79 3E−10 215315_at zinc finger protein 549 ZNF549 1.79 5E−05 242517_at G protein-coupled receptor 54 GPR54 1.79 8E−13 224062_x_at “kallikrein 4 (prostase, enamel matrix, prostate)” KLK4 1.79 1E−05 207686_s_at “caspase 8, apoptosis-related cysteine protease” CASP8 1.79 5E−08 231794_at cytotoxic T-lymphocyte-associated protein 4 CTLA4 1.79 7E−08 1560686_at “integrin, alpha D” ITGAD 1.78 5E−11 1562338_at Membrane-associated RING-CH protein I MARCH-I 1.78 2E−05 227690_at “Gamma-aminobutyric acid (GABA) A receptor, beta 3” GABRB3 1.78 2E−05 224360_s_at protein kinase C and casein kinase substrate in neurons 1 | PACSIN1 1.78 2E−05 protein kinase C and casein kinase substrate in neurons 1 213834_at IQ motif and Sec7 domain 3 IQSEC3 1.78 2E−06 205699_at mitogen-activated protein kinase kinase 6 MAP2K6 1.78 5E−08 221558_s_at lymphoid enhancer-binding factor 1 LEF1 1.78 2E−07 235768_at SH3 domain containing ring finger 2 SH3RF2 1.77 3E−05 224367_at brain expressed X-linked 2 | brain expressed X-linked 2 BEX2 1.77 6E−08 220369_at KIAA2010 KIAA2010 1.77 8E−06 224204_x_at aryl hydrocarbon receptor nuclear translocator-like 2 ARNTL2 1.77 9E−07 209911_x_at “histone 1, H2bd” HIST1H2BD 1.77 3E−13 211085_s_at serine/threonine kinase 4 | serine/threonine kinase 4 STK4 1.77 3E−12 215332_s_at “CD8 antigen, beta polypeptide 1 (p37)” CD8B1 1.77 7E−05 222927_s_at “lectin, mannose-binding, 1 like” LMAN1L 1.77 3E−05 206316_s_at kinetochore associated 1 KNTC1 1.77 7E−14 1555638_a_at “SAM domain, SH3 domain and nuclear localisation signals, 1” SAMSN1 1.77 1E−11 232372_at Tubby like protein 4 TULP4 1.77 2E−06 211003_x_at “transglutaminase 2 (C polypeptide, protein-glutamine- TGM2 1.77 1E−06 gamma-glutamyltransferase)” 214217_at “Glutamate receptor, metabotropic 5” GRM5 1.77 3E−06 221251_x_at high mobility group AT-hook 1-like 4 | high mobility group HMGA1L4 1.77 4E−06 AT-hook 1-like 4 222943_at “glucosidase, beta, acid 3 (cytosolic)” GBA3 1.77 2E−09 235104_at leukocyte-derived arginine aminopeptidase LRAP 1.77 1E−05 208261_x_at “interferon, alpha 10” IFNA10 1.76 1E−06 204205_at “apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3G” APOBEC3G 1.76 4E−09 1565483_at “epidermal growth factor receptor (erythroblastic leukemia viral EGFR 1.76 8E−05 (v-erb-b) oncogene homolog, avian)” 1557924_s_at “alkaline phosphatase, liver/bone/kidney” ALPL 1.76 2E−05 206999_at “interleukin 12 receptor, beta 2” IL12RB2 1.76 1E−05 215573_at Catalase CAT 1.76 5E−08 208537_at “endothelial differentiation, sphingolipid G-protein-coupled EDG5 1.76 4E−08 receptor, 5” 214539_at “serine (or cysteine) proteinase inhibitor, clade B SERPINB10 1.76 1E−06 (ovalbumin), member 10” 1569909_at keratin 6L KRT6L 1.75 3E−09 206720_at “mannosyl (alpha-1,6-)-glycoprotein beta-1,6-N-acetyl- MGAT5 1.75 3E−05 glucosaminyltransferase” 241049_at “glutamate receptor, metabotropic 7” GRM7 1.75 6E−09 1553552_at trace amine receptor 5 TRAR5 1.75 4E−07 222774_s_at neuropilin (NRP) and tolloid (TLL)-like 2 NETO2 1.75 8E−06 217407_x_at peptidylprolyl isomerase (cyclophilin)-like 2 PPIL2 1.75 2E−05 204935_at “protein tyrosine phosphatase, non-receptor type 2” PTPN2 1.75 7E−10 230717_at lipocalcin 12 LCN12 1.75 1E−05 1565935_at prematurely terminated mRNA decay factor-like LOC91431 1.75 4E−05 1555048_a_at chromosome 21 open reading frame 29 C21orf29 1.75 7E−05 233362_at zinc finger protein 341 ZNF341 1.75 2E−06 227677_at “Janus kinase 3 (a protein tyrosine kinase, leukocyte)” JAK3 1.74 1E−08 1556282_at FGFR1 oncogene partner 2 FGFR1OP2 1.74 3E−06 206901_at hypothetical protein MGC11271 MGC11271 1.74 5E−05 1562590_at hypothetical protein FLJ25756 FLJ25756 1.74 6E−06 220330_s_at “SAM domain, SH3 domain and nuclear localisation signals, 1” SAMSN1 1.74 3E−12 1555277_a_at “solute carrier family 4, sodium bicarbonate cotransporter, member 5” SLC4A5 1.74 6E−06 1554300_a_at hypothetical protein LOC136306 LOC136306 1.74 5E−05 222257_s_at angiotensin I converting enzyme (peptidyl-dipeptidase A) 2 ACE2 1.74 1E−05 1570516_s_at “olfactory receptor, family 51, subfamily B, member 5” OR51B5 1.74 5E−07 231332_at Hypothetical protein LOC196394 LOC196394 1.73 6E−08 208259_x_at “interferon, alpha 7” IFNA7 1.73 2E−06 211424_x_at NA NA 1.73 9E−06 205552_s_at “2′,5′-oligoadenylate synthetase 1, 40/46 kDa” OAS1 1.73 5E−05 204056_s_at mevalonate kinase (mevalonic aciduria) MVK 1.73 2E−06 232562_at “CDNA FLJ11554 fis, clone HEMBA1003037” NA 1.73 3E−05 239419_at “Protein tyrosine phosphatase, receptor type, A” VPS16 1.73 1E−07 1552504_a_at BR serine/threonine kinase 1 BRSK1 1.73 4E−10 213472_at heterogeneous nuclear ribonucleoprotein H1 (H) HNRPH1 1.73 4E−06 207854_at glycophorin E GYPE 1.73 3E−06 224555_x_at “interleukin 1 family, member 7 (zeta)” IL1F7 1.73 5E−09 215442_s_at thyroid stimulating hormone receptor TSHR 1.73 8E−05 206429_at coagulation factor II (thrombin) receptor-like 1 F2RL1 1.73 4E−05 207441_at submaxillary gland androgen regulated protein 3 homolog B (mouse) SMR3B 1.73 4E−05 239646_at KIAA1961 protein RAPGEF6 1.73 7E−07 1552506_at crumbs homolog 2 (Drosophila) CRB2 1.73 1E−05 219259_at “sema domain, immunoglobulin domain (Ig), transmembrane SEMA4A 1.73 8E−10 domain (TM) and short cytoplasmic domain, (semaphorin) 4A” 232722_at ribonuclease T2 RNASET2 1.73 4E−08 207957_s_at “Protein kinase C, beta 1” PRKCB1 1.72 2E−05 1569830_at “Protein tyrosine phosphatase, receptor type, C” PTPRC 1.72 9E−06 1554208_at similar to mouse meiosis defective 1 gene MGC40042 1.72 2E−15 238474_at nucleoporin 43 kDa NUP43 1.72 3E−07 220315_at “poly (ADP-ribose) polymerase family, member 11” PARP11 1.72 9E−07 216407_at Vac14 homolog (S. cerevisiae) VAC14 1.72 1E−06 238853_at Glioma amplified sequence 64 NA 1.72 9E−06 211360_s_at “inositol 1,4,5-triphosphate receptor, type 2” ITPR2 1.71 1E−08 1561578_s_at Similar to mitochondrial carrier triple repeat 1 NA 1.71 4E−06 238699_s_at calcium/calmodulin-dependent serine protein kinase (MAGUK family) CASK 1.71 3E−06 235650_at hypothetical protein FLJ23834 FLJ23834 1.71 9E−09 240703_s_at hect (homologous to the E6-AP (UBE3A) carboxyl terminus) HERC1 1.71 4E−05 domain and RCC1 (CHC1)-like domain (RLD) 1 221696_s_at serine/threonine/tyrosine kinase 1 | serine/threonine/tyrosine kinase 1 STYK1 1.71 3E−08 1552742_at “potassium voltage-gated channel, subfamily H (eag-related), member 8” KCNH8 1.71 4E−05 223809_at regulator of G-protein signalling 18 RGS18 1.71 2E−11 241737_x_at Vaccinia related kinase 1 VRK1 1.71 3E−07 242077_x_at chromosome 6 open reading frame 150 C6orf150 1.71 6E−09 221004_s_at integral membrane protein 2C | integral membrane protein 2C ITM2C 1.71 3E−06 1552343_s_at phosphodiesterase 7A PDE7A 1.71 6E−07 1552801_at calpain 13 CAPN13 1.70 4E−06 206618_at interleukin 18 receptor 1 IL18R1 1.70 1E−04 205291_at “interleukin 2 receptor, beta | interleukin 2 receptor, beta” IL2RB 1.70 1E−10 207976_at kelch-like 18 (Drosophila) KLHL18 1.70 7E−09 1556412_at KIAA0478 gene product KIAA0478 1.70 1E−06 205269_at lymphocyte cytosolic protein 2 (SH2 domain containing LCP2 1.70 1E−09 leukocyte protein of 76 kDa) 205016_at “transforming growth factor, alpha” TGFA 1.70 8E−09 1560803_at “dynein, axonemal, heavy polypeptide 3” DNAH3 1.70 1E−05 235061_at protein phosphatase 1K (PP2C domain containing) PPM1K 1.70 3E−12 1552857_a_at 5-hydroxytryptamine (serotonin) receptor 6 HTR6 1.70 2E−05 1563744_a_at Disrupted in schizophrenia 1 DISC1 1.69 4E−05 241357_at extracellular signal-regulated kinase 8 ERK8 1.69 3E−06 237036_at F-box protein 10 FBXO10 1.69 4E−05 221293_s_at differentially expressed in FDCP 6 homolog (mouse) DEF6 1.69 2E−10 235169_at F-box protein 27 FBXO27 1.69 2E−05 211207_s_at acyl-CoA synthetase long-chain family member 6 ACSL6 1.69 8E−06 1559425_at “Protein kinase C, eta” PRKCH 1.69 8E−07 205668_at lymphocyte antigen 75 LY75 1.69 1E−07 1566963_at NA NA 1.69 2E−05 208267_at “transient receptor potential cation channel, subfamily V, member 5” TRPV5 1.69 2E−08 210670_at pancreatic polypeptide PPY 1.69 3E−09 220422_at ubiquilin 3 UBQLN3 1.69 7E−09 1555671_at “amyotrophic lateral sclerosis 2 (juvenile) chromosome ALS2CR14 1.69 9E−08 region, candidate 14” 244692_at hypothetical protein FLJ39501 FLJ39501 1.69 8E−05 1559159_at KIAA0582 KIAA0582 1.69 5E−10 209006_s_at NPD014 protein NPD014 1.68 2E−08 219391_at “3′(2′),5′-bisphosphate nucleotidase 1” BPNT1 1.68 6E−09 215470_at “CDNA FLJ36630 fis, clone TRACH2018278” NA 1.68 2E−05 208481_at ankyrin repeat and SOCS box-containing 4 ASB4 1.68 2E−05 240770_at hypothetical protein LOC134285 LOC134285 1.68 5E−05 219285_s_at ninein (GSK3B interacting protein) NIN 1.68 1E−05 227152_at hypothetical protein FLJ20696 FLJ20696 1.68 4E−12 211080_s_at NIMA (never in mitosis gene a)-related kinase 2 | NEK2 1.68 2E−06 NIMA (never in mitosis gene a)-related kinase 2 207735_at ring finger protein 125 RNF125 1.68 6E−10 228288_at NA NA 1.68 2E−07 223605_at “solute carrier family 25 (mitochondrial carrier), member 18” SLC25A18 1.67 6E−06 230036_at chromosome 7 open reading frame 6 C7orf6 1.67 1E−10 1553177_at SH2 domain-containing molecule EAT2 EAT2 1.67 9E−09 237407_at HS1-binding protein 3 FLJ14249 1.67 5E−05 207509_s_at leukocyte-associated Ig-like receptor 2 LAIR2 1.67 7E−05 227811_at “FGD1 family, member 3” FGD3 1.67 2E−09 240843_at “Protein tyrosine phosphatase, non-receptor type 2” PTPN2 1.67 6E−06 217495_x_at “calcitonin/calcitonin-related polypeptide, alpha” CALCA 1.67 1E−05 241988_x_at Slingshot homolog 2 (Drosophila) SSH2 1.67 2E−07 207162_s_at “calcium channel, voltage-dependent, L type, alpha 1B subunit” CACNA1B 1.67 3E−09 220214_at zinc finger protein 215 ZNF215 1.67 2E−05 1563590_at “Protein tyrosine phosphatase, receptor type, E” PTPRE 1.67 1E−06 207106_s_at leukocyte tyrosine kinase LTK 1.66 1E−07 207114_at “lymphocyte antigen 6 complex, locus G6C” LY6G6C 1.66 1E−07 219734_at “SID1 transmembrane family, member 1” SIDT1 1.66 2E−10 205255_x_at “transcription factor 7 (T-cell specific, HMG-box)” TCF7 1.66 2E−08 206916_x_at tyrosine aminotransferase TAT 1.66 8E−06 222746_s_at B-box and SPRY domain containing BSPRY 1.66 1E−13 220420_at “lectin, mannose-binding, 1 like” LMAN1L 1.66 7E−06 234092_s_at transmembrane 6 superfamily member 2 TM6SF2 1.66 6E−08 222940_at “sulfotransferase family 1E, estrogen-preferring, member 1” SULT1E1 1.66 7E−05 226906_s_at Rho GTPase activating protein 9 ARHGAP9 1.66 2E−09 224290_at NA NA 1.66 6E−06 219695_at “sphingomyelin phosphodiesterase 3, neutral membrane SMPD3 1.66 3E−08 (neutral sphingomyelinase II)” 244011_at protein phosphatase 1K (PP2C domain containing) PPM1K 1.66 6E−10 227609_at epithelial stromal interaction 1 (breast) EPSTI1 1.66 9E−07 1552703_s_at “caspase 1, apoptosis-related cysteine protease (interleukin 1, CASP1 | 1.66 4E−12 beta, convertase) | CARD only protein” COPI 232383_at Transcription factor EC TFEC 1.66 3E−07 1553823_a_at receptor transporting protein 1 RTP1 1.66 3E−05 221340_at caudal type homeo box transcription factor 4 CDX4 1.65 7E−06 227344_at “Zinc finger protein, subfamily 1A, 1 (Ikaros)” ZNFN1A1 1.65 2E−11 229312_s_at G kinase anchoring protein 1 GKAP1 1.65 2E−09 203148_s_at tripartite motif-containing 14 TRIM14 1.65 4E−07 227552_at septin 1 1-Sep 1.65 4E−08 227462_at Leukocyte-derived arginine aminopeptidase LRAP 1.65 2E−05 1553666_at NY-REN-41 antigen NY-REN-41 1.65 8E−05 239885_at Transcribed locus NA 1.65 2E−09 235720_at cysteine-rich protein 3 CRIP3 1.65 1E−05 208286_x_at “POU domain, class 5, transcription factor 1” POU5F1 1.65 1E−07 213109_at NA NA 1.65 8E−11 205299_s_at “butyrophilin, subfamily 2, member A2” BTN2A2 1.65 5E−10 236002_at TAO kinase 3 TAOK3 1.64 8E−09 1565818_s_at “zinc finger protein, subfamily 1A, 1 (Ikaros)” ZNFN1A1 1.64 1E−05 203110_at PTK2B protein tyrosine kinase 2 beta PTK2B 1.64 2E−08 207592_s_at hyperpolarization activated cyclic nucleotide-gated HCN2 1.64 3E−06 potassium channel 2 223746_at serine/threonine kinase 4 STK4 1.64 9E−08 239438_at Rap guanine nucleotide exchange factor (GEF) 6 RAPGEF6 1.64 6E−05 240189_at Transcribed locus NA 1.64 9E−08 220440_at “lectin, galactoside-binding, soluble, 13 (galectin 13)” LGALS13 1.64 2E−05 205538_at “coronin, actin binding protein, 2A” CORO2A 1.64 1E−08 216997_x_at “transducin-like enhancer of split 4 (E(sp1) homolog, Drosophila)” TLE4 1.64 5E−09 236305_at LOC317671 LOC317671 1.64 3E−09 209685_s_at “protein kinase C, beta 1” PRKCB1 1.64 5E−06 1564231_at KIAA1374 protein KIAA1374 1.64 5E−10 236236_at “CDNA FLJ30437 fis, clone BRACE2009045” NA 1.64 8E−05 206974_at chemokine (C-X-C motif) receptor 6 CXCR6 1.64 4E−07 242175_at X-ray radiation resistance associated 1 XRRA1 1.64 3E−05 207990_x_at acrosomal vesicle protein 1 ACRV1 1.64 8E−11 216377_x_at “alkaline phosphatase, placental-like 2” ALPPL2 1.63 2E−06 243410_at “Protein tyrosine phosphatase, non-receptor type 2” PTPN2 1.63 9E−08 214624_at uroplakin 1A UPK1A 1.63 1E−07 206248_at “protein kinase C, epsilon” PRKCE 1.63 4E−07 216112_at Protein kinase N2 PKN2 1.63 2E−06 208203_x_at “killer cell immunoglobulin-like receptor, two domains, KIR2DS5 1.63 2E−08 short cytoplasmic tail, 5” 210969_at protein kinase N2 PKN2 1.63 4E−08 207838_x_at pre-B-cell leukemia transcription factor interacting protein 1 PBXIP1 1.63 1E−07 209569_x_at DNA segment on chromosome 4 (unique) 234 expressed sequence D4S234E 1.63 8E−10 238054_at ADP-ribosylhydrolase like 1 ADPRHL1 1.63 4E−06 207382_at tumor protein p73-like TP73L 1.63 6E−05 216624_s_at “myeloid/lymphoid or mixed-lineage leukemia MLL 1.63 3E−05 (trithorax homolog, Drosophila)” 217566_s_at transglutaminase 4 (prostate) TGM4 1.63 8E−05 223423_at G protein-coupled receptor 160 GPR160 1.63 8E−09 213017_at abhydrolase domain containing 3 ABHD3 1.63 4E−11 227736_at chromosome 10 open reading frame 99 C10orf99 1.63 7E−05 1555581_a_at tumor protein p73-like TP73L 1.62 6E−05 219411_at “engulfment and cell motility 3 (ced-12 homolog, C. elegans)” ELMO3 1.62 2E−09 206830_at “solute carrier family 4, sodium bicarbonate transporter-like, SLC4A10 1.62 3E−05 member 10” 211598_x_at vasoactive intestinal peptide receptor 2 | vasoactive intestinal VIPR2 | 1.62 9E−06 peptide receptor 2 | similar to vasoactive intestinal peptide LOC441305 receptor 2 | similar to vasoactive intestinal peptide receptor 2 210743_s_at CDC14 cell division cycle 14 homolog A (S. cerevisiae) CDC14A 1.62 6E−11 232995_at DNA-damage inducible protein 1 PDGFD 1.62 2E−05 231230_at Clone 23786 mRNA sequence NA 1.62 8E−05 244394_at B lymphoid tyrosine kinase BLK 1.62 8E−05 211888_x_at “caspase 10, apoptosis-related cysteine protease” CASP10 1.62 1E−04 216522_at “olfactory receptor, family 2, subfamily B, member 6” OR2B6 1.62 2E−08 241913_at Hypothetical gene supported by AK123781 NA 1.62 1E−06 204044_at quinolinate phosphoribosyltransferase (nicotinate-nucleotide QPRT 1.62 1E−08 pyrophosphorylase (carboxylating)) 206835_at statherin STATH 1.62 5E−05 234775_at “olfactory receptor, family 51, subfamily B, member 5” OR51B5 1.62 7E−10 211005_at linker for activation of T cells LAT 1.62 3E−08 204882_at NA NA 1.62 1E−07 205639_at acyloxyacyl hydrolase (neutrophil) AOAH 1.62 2E−06 1553102_a_at DKFZP434C171 protein DKFZP434C171 1.62 1E−12 223607_x_at “zinc finger, SWIM domain containing 1” ZSWIM1 1.62 2E−08 226313_at chromosome 10 open reading frame 35 C10orf35 1.61 3E−09 217711_at “TEK tyrosine kinase, endothelial (venous malformations, TEK 1.61 6E−05 multiple cutaneous and mucosal)” 225931_s_at chromosome 17 open reading frame 27 C17orf27 1.61 9E−11 204798_at v-myb myeloblastosis viral oncogene homolog (avian) MYB 1.61 2E−06 204994_at myxovirus (influenza virus) resistance 2 (mouse) MX2 1.61 7E−06 49049_at deltex 3 homolog (Drosophila) DTX3 1.61 1E−06 215170_s_at KIAA0912 protein Cep152 1.61 7E−08 216288_at cysteinyl leukotriene receptor 1 CYSLTR1 1.61 4E−06 1552915_at “interleukin 28A (interferon, lambda 2)” IL28A 1.61 4E−09 242976_at “Homo sapiens, clone IMAGE: 5286757, mRNA” NA 1.61 2E−05 210629_x_at leukocyte specific transcript 1 LST1 1.61 4E−08 228551_at hypothetical protein MGC24039 MGC24039 1.61 2E−09 205509_at carboxypeptidase B1 (tissue) CPB1 1.61 3E−08 206295_at interleukin 18 (interferon-gamma-inducing factor) IL18 1.61 2E−07 1557193_at “Protein tyrosine phosphatase, non-receptor type 2” PTPN2 1.61 1E−09 219045_at “ras homolog gene family, member F (in filopodia)” RHOF 1.61 6E−07 213990_s_at p21(CDKN1A)-activated kinase 7 PAK7 1.61 4E−07 234637_at keratin associated protein 4-5 KRTAP4-5 1.60 7E−05 206663_at Sp4 transcription factor SP4 1.60 2E−06 221578_at Ras association (RaIGDS/AF-6) domain family 4 RASSF4 1.60 6E−05 232721_at tripartite motif-containing 55 TRIM55 1.60 7E−05 1552796_a_at single-minded homolog 1 (Drosophila) SIM1 1.60 1E−05 210742_at CDC14 cell division cycle 14 homolog A (S. cerevisiae) CDC14A 1.60 9E−07 1569257_at formin-like 1 FMNL1 1.60 5E−06 207217_s_at NADPH oxidase 1 NOX1 1.60 1E−06 1562238_at Chromosome 13 open reading frame 22 C13orf22 1.60 8E−07 1555248_a_at WNK lysine deficient protein kinase 3 WNK3 1.60 2E−05 216341_s_at gonadotropin-releasing hormone receptor GNRHR 1.60 4E−07 1553556_at “taste receptor, type 2, member 40” TAS2R40 1.60 1E−05 1565635_at NA NA 1.60 2E−05 240446_at Mindbomb homolog 1 (Drosophila) MIB1 1.60 3E−05 242549_at protein kinase D3 PRKD3 1.60 3E−09 205411_at serine/threonine kinase 4 STK4 1.60 1E−09 217701_x_at NA NA 1.60 6E−05 239529_at dendritic cell nuclear protein 1 DCNP1 1.60 9E−05 215275_at TRAF3-interacting Jun N-terminal kinase (JNK)-activating modulator T3JAM 1.60 2E−07 1565674_at “Fc fragment of IgG, low affinity IIa, receptor (CD32)” FCGR2A 1.60 1E−04 234329_at chloride intracellular channel 5 CLIC5 1.60 1E−05 213778_x_at zinc finger protein 276 homolog (mouse) ZFP276 1.60 1E−10 211834_s_at tumor protein p73-like TP73L 1.60 9E−05 220252_x_at chromosome X open reading frame 21 CXorf21 1.59 8E−10 1555423_at slingshot homolog 2 (Drosophila) SSH2 1.59 8E−05 219977_at aryl hydrocarbon receptor interacting protein-like 1 AIPL1 1.59 3E−07 203317_at pleckstrin and Sec7 domain containing 4 PSD4 1.59 9E−09 206280_at “cadherin 18, type 2” CDH18 1.59 2E−05 213953_at keratin 20 KRT20 1.59 9E−05 1556925_at Chondroitin sulfate proteoglycan 6 (bamacan) CSPG6 1.59 4E−09 220374_at BTB (POZ) domain containing 5 BTBD5 1.59 3E−07 227496_at hypothetical protein LOC253842 LOC253842 1.59 4E−07 227817_at “Protein kinase C, beta 1” PRKCB1 1.59 1E−05 211117_x_at estrogen receptor 2 (ER beta) ESR2 1.59 3E−05 203930_s_at microtubule-associated protein tau MAPT 1.59 7E−05 236296_x_at NA NA 1.59 9E−05 1553426_at hypothetical protein FLJ37543 FLJ37543 1.59 3E−07 235971_at TRAF-interacting protein with a forkhead-associated domain TIFA 1.59 3E−09 241736_at F-box and WD-40 domain protein 2 FBXW2 1.59 6E−08 210359_at metastasis suppressor 1 MTSS1 1.58 1E−07 205599_at TNF receptor-associated factor 1 TRAF1 1.58 1E−11 221999_at vaccinia related kinase 3 VRK3 1.58 8E−13 220702_at NA NA 1.58 3E−08 1555651_at “olfactory receptor, family 10, subfamily A, member 5” OR10A5 1.58 7E−05 216621_at NA NA 1.58 4E−06 214339_s_at mitogen-activated protein kinase kinase kinase kinase 1 MAP4K1 1.58 2E−06 234455_at zinc finger protein 1 homolog (mouse) ZFP1 1.58 9E−05 212588_at “protein tyrosine phosphatase, receptor type, C” PTPRC 1.58 1E−06 1567008_at NA NA 1.58 3E−05 231874_at NA NA 1.58 3E−10 231263_at chromosome 6 open reading frame 81 C6orf81 1.58 2E−05 211778_s_at zinc finger protein 339 | zinc finger protein 339 ZNF339 1.58 7E−07 206372_at myogenic factor 6 (herculin) MYF6 1.58 6E−05 1564029_at ubiquitin specific protease 49 MGC20741 1.58 5E−05 219233_s_at gasdermin-like GSDML 1.58 3E−05 1553373_at NA NA 1.58 1E−05 227607_at associated molecule with the SH3 domain of STAM (AMSH) AMSH-LP 1.58 1E-10 like protein 1562307_at Ring finger protein 24 RNF24 1.58 7E−06 221551_x_at “ST6 (alpha-N-acetyl-neuraminy1-2,3-beta-galactosy1-1,3)-N- ST6GALNAC4 1.58 1E−08 acetylgalactosaminide alpha-2,6-sialyltransferase 4” 206674_at fms-related tyrosine kinase 3 FLT3 1.58 1E−05 207725_at “POU domain, class 4, transcription factor 2” POU4F2 1.58 2E−05 235498_at hypothetical protein MGC22773 MGC22773 1.58 7E−05 1558603_at plasminogen-like PLGL 1.57 3E−05 1555035_a_at Usher syndrome 3A USH3A 1.57 6E−05 217603_at “ATPase, H+ transporting, lysosomal V0 subunit a isoform 2” ATP6V0A2 1.57 8E−06 207859_s_at “cholinergic receptor, nicotinic, beta polypeptide 3” CHRNB3 1.57 2E−08 1559624_at Serine/threonine kinase 32A STK32A 1.57 5E−05 242765_at Myelin-associated oligodendrocyte basic protein MOBP 1.57 1E−05 239955_at Transcribed locus NA 1.57 5E−06 209604_s_at GATA binding protein 3 GATA3 1.57 6E−06 218792_s_at B-box and SPRY domain containing BSPRY 1.57 7E−06 210865_at “Fas ligand (TNF superfamily, member 6)” FASLG 1.57 9E−09 223245_at spermatid perinuclear RNA binding protein STRBP 1.57 3E−14 215512_at membrane-associated ring finger (C3HC4) 6 6-Mar 1.57 6E−07 1558561_at histocompatibility (minor) 13 HM13 1.57 6E−08 207759_s_at disrupted in schizophrenia 1 DISC1 1.57 1E−08 1569462_x_at “potassium channel, subfamily T, member 1” KCNT1 1.57 3E−05 230983_at B-cell novel protein 1 BCNP1 1.57 1E−08 242130_at NA NA 1.57 6E−06 230262_at “CDNA FLJ30377 fis, clone BRACE2007952” NA 1.57 6E−07 238210_at RYK receptor-like tyrosine kinase RYK 1.56 4E−06 232024_at “GTPase, IMAP family member 2” GIMAP2 1.56 2E−10 1555785_a_at 5′-3′ exoribonuclease 1 XRN1 1.56 9E−09 207164_s_at zinc finger protein 238 ZNF238 1.56 7E−06 230217_at hypothetical protein MGC34646 MGC34646 1.56 6E−06 213631_x_at “CDNA FLJ40920 fis, clone UTERU2005905” NA 1.56 5E−10 230499_at Baculoviral IAP repeat-containing 3 BIRC3 1.56 6E−08 211226_at galanin receptor 2 GALR2 1.56 9E−05 219209_at interferon induced with helicase C domain 1 IFIH1 1.56 1E−06 1561206_at kelch-like 8 (Drosophila) KLHL8 1.56 2E−07 1560665_at serine carboxypeptidase 1 SCPEP1 1.56 2E−06 1553120_at claspin homolog (Xenopus laevis) CLSPN 1.56 6E−05 1554253_a_at LAG1 longevity assurance homolog 3 (S. cerevisiae) LASS3 1.56 5E−08 244565_at Similar to Hmx2 protein NA 1.56 5E−05 231291_at Gastric inhibitory polypeptide receptor GIPR 1.56 5E−05 38241_at “butyrophilin, subfamily 3, member A3” BTN3A3 1.56 1E−10 1552491_at isopentenyl-diphosphate delta isomerase 2 IDI2 1.56 2E−05 226070_at hypothetical protein LOC286257 LOC286257 1.56 2E−11 220684_at T-box 21 TBX21 1.56 1E−10 222812_s_at “ras homolog gene family, member F (in filopodia)” RHOF 1.56 4E−09 206249_at mitogen-activated protein kinase kinase kinase 13 MAP3K13 1.55 9E−05 1569540_at NA NA 1.55 6E−06 236281_x_at 5-hydroxytryptamine (serotonin) receptor 7 (adenylate cyclase-coupled) HTR7 1.55 8E−05 1564757_a_at hypothetical protein BC015395 LOC130940 1.55 4E−05 224046_s_at phosphodiesterase 7A PDE7A 1.55 4E−09 229168_at “collagen, type XXIII, alpha 1” COL23A1 1.55 3E−08 229419_at “F-box and WD-40 domain protein 7 (archipelago homolog, Drosophila)” FBXW7 1.55 1E−11 221080_s_at “family with sequence similarity 31, member C” FAM31C 1.55 1E−09 221339_at “olfactory receptor, family 10, subfamily C, member 1” OR10C1 1.55 2E−08 219066_at hypothetical protein MDS018 MDS018 1.55 6E−13 205101_at MHC class II transactivator MHC2TA 1.55 3E−05 209703_x_at DKFZP586A0522 protein DKFZP586A0522 1.55 6E−07 1553681_a_at perforin 1 (pore forming protein) PRF1 1.55 2E−13 219786_at “metallothionein-like 5, testis-specific (tesmin)” MTL5 1.55 1E−07 243764_at V-set and immunoglobulin domain containing 1 VSIG1 1.55 2E−06 231112_at Small nuclear ribonucleoprotein polypeptide E SNRPE 1.55 6E−07 227002_at chromosome 9 open reading frame 59 C9orf59 1.55 5E−09 1557145_at Serine/threonine kinase 38 STK38 1.55 1E−06 242211_x_at hypothetical protein KIAA1924 KIAA1924 1.55 3E−07 209007_s_at NPD014 protein NPD014 1.55 2E−12 218811_at chromosome 7 open reading frame 19 C7orf19 1.55 1E−09 243430_at seizure related 6 homolog (mouse) SEZ6 1.55 4E−05 213947_s_at nucleoporin 210 kDa NUP210 1.55 7E−06 231599_x_at “D4, zinc and double PHD fingers family 1” DPF1 1.55 3E−05 206776_x_at acrosomal vesicle protein 1 ACRV1 1.54 5E−10 211531_x_at proline-rich protein BstNI subfamily 1 PRB1 1.54 6E−05 233343_at NA NA 1.54 2E−07 219683_at frizzled homolog 3 (Drosophila) FZD3 1.54 6E−07 204950_at “caspase recruitment domain family, member 8” CARD8 1.54 4E−13 217291_at NA NA 1.54 7E−05 226433_at ring finger protein 157 RNF157 1.54 1E−06 209863_s_at tumor protein p73-like TP73L 1.54 2E−09 205739_x_at Zinc finger protein (ZFD25) ZFD25 1.54 2E−10 239149_at Bromodomain containing 4 BRD4 1.54 2E−05 206261_at zinc finger protein 239 ZNF239 1.54 3E−10 231796_at EPH receptor A8 EPHA8 1.54 1E−07 211828_s_at TRAF2 and NCK interacting kinase TNIK 1.54 1E−08 210934_at B lymphoid tyrosine kinase BLK 1.54 2E−08 1565131_x_at Mitogen-activated protein kinase kinase kinase 2 MAP3K2 1.54 3E−05 208341_x_at chorionic somatomammotropin hormone 2 CSH2 1.54 5E−08 211573_x_at “transglutaminase 2 (C polypeptide, protein-glutamine- TGM2 1.54 4E−05 gamma-glutamyltransferase)” 1569614_s_at lipocalin 8 LCN8 1.54 9E−05 205659_at histone deacetylase 9 HDAC9 1.54 1E−06 1552788_a_at helicase (DNA) B HELB 1.54 5E−05 205801_s_at RAS guanyl releasing protein 3 (calcium and DAG-regulated) RASGRP3 1.54 2E−06 1552555_at polyserase-2 FLJ90661 1.54 4E−07 1565588_at SP140 nuclear body protein SP140 1.54 2E−06 213756_s_at heat shock transcription factor 1 HSF1 1.54 5E−09 1562026_at WNK lysine deficient protein kinase 2 WNK2 1.53 2E−06 220987_s_at chromosome 11 open reading frame 17 | chromosome 11 open reading C11orf17 | 1.53 2E−08 frame 17 | likely ortholog of rat SNF1/AMP-activated protein kinase | SNARK likely ortholog of rat SNF1/AMP-activated protein kinase 210894_s_at centrosomal protein 2 CEP2 1.53 8E−06 205009_at “trefoil factor 1 (breast cancer, estrogen-inducible sequence expressed in)” TFF1 1.53 3E−05 222951_s_at ankyrin repeat domain 5 ANKRD5 1.53 3E−05 211333_s_at “Fas ligand (TNF superfamily, member 6)” FASLG 1.53 3E−08 215086_at Inhibitor of Bruton agammaglobulinemia tyrosine kinase IBTK 1.53 6E−05 221209_s_at otoraplin OTOR 1.53 6E−05 205733_at Bloom syndrome BLM 1.53 4E−11 224156_x_at interleukin 17 receptor B IL17RB 1.53 2E−08 202558_s_at “stress 70 protein chaperone, microsome-associated, 60 kDa” STCH 1.53 1E−07 211368_s_at “caspase 1, apoptosis-related cysteine protease CASP1 1.53 6E−10 (interleukin 1, beta, convertase)” 226474_at nucleotide-binding oligomerization domains 27 NOD27 1.53 3E−08 206353_at cytochrome c oxidase subunit VIa polypeptide 2 COX6A2 1.53 7E−07 211516_at “interleukin 5 receptor, alpha” IL5RA 1.53 4E−06 236240_at hypothetical protein FLJ21106 FLJ21106 1.53 2E−07 224451_x_at Rho GTPase activating protein 9 | Rho GTPase activating protein 9 ARHGAP9 1.53 1E−07 219112_at Rap guanine nucleotide exchange factor (GEF) 6 RAPGEF6 1.53 1E−10 231962_at “adaptor-related protein complex 4, beta 1 subunit” AP4B1 1.53 2E−07 207408_at “solute carrier family 22 (organic cation transporter), member 14” SLC22A14 1.53 2E−10 233575_s_at “transducin-like enhancer of split 4 (E(sp1) homolog, Drosophila)” TLE4 1.53 2E−06 1553549_at vomeronasal 1 receptor 2 VN1R2 1.53 5E−05 1554116_s_at “poly (ADP-ribose) polymerase family, member 11” PARP11 1.53 9E−06 1558755_x_at hypothetical protein LOC284390 LOC284390 1.53 3E−10 207359_at “calcium/calmodulin-dependent protein kinase kinase 2, beta” CAMKK2 1.53 8E−05 211194_s_at tumor protein p73-like TP73L 1.53 5E−05 220958_at Hypothetical protein FLJ20574 FLJ20574 1.53 5E−06 210038_at “protein kinase C, theta” PRKCQ 1.52 4E−09 224209_s_at guanine deaminase GDA 1.52 9E−05 234943_at “casein kinase 1, epsilon | TPTE/TPIP pseudogene | similar to TPTE and CSNK1E | 1.52 5E−07 PTEN homologous inositol lipid phosphatase isoform alpha; TPTE and LOC387593 | PTEN homologous inositol lipid phosphatase” LOC400927 225795_at hypothetical gene supported by AL449243 LOC91689 1.52 1E−08 242968_at NA NA 1.52 1E−07 242970_at KIAA1463 protein KIAA1463 1.52 3E−06 210234_at “glutamate receptor, metabotropic 4” GRM4 1.52 1E−07 241920_x_at hypothetical protein FLJ21439 FLJ21439 1.52 9E−10 231612_at testis development protein NYD-SP26 NYD-SP26 1.52 8E−05 230756_at hypothetical protein MGC33414 MGC33414 1.52 6E−06 239554_at Ring finger protein 13 RNF13 1.52 5E−05 214791_at hypothetical protein BC004921 LOC93349 1.52 2E−12 234820_at MAS1 oncogene-like MAS1L 1.52 2E−05 205235_s_at M-phase phosphoprotein 1 MPHOSPH1 1.52 6E−06 220989_s_at amnionless homolog (mouse) | amnionless homolog (mouse) AMN 1.52 1E−04 226525_at Serine/threonine kinase 17b (apoptosis-inducing) STK17B 1.52 3E−11 205586_x_at VGF nerve growth factor inducible VGF 1.52 8E−06 1570231_at “LATS, large tumor suppressor, homolog 1 (Drosophila)” LATS1 1.52 5E−05 226117_at TRAF-interacting protein with a forkhead-associated domain TIFA 1.52 2E−14 214572_s_at insulin-like 3 (Leydig cell) INSL3 1.52 7E−05 211044_at tripartite motif-containing 14 | tripartite motif-containing 14 TRIM14 1.52 3E−06 228607_at “2′-5′-oligoadenylate synthetase 2, 69/71 kDa” OAS2 1.52 9E−07 1558754_at hypothetical protein LOC284390 LOC284390 1.52 2E−06 220390_at hypothetical protein FLJ23598 FLJ23598 1.52 9E−05 219413_at acyl-Coenzyme A binding domain containing 4 ACBD4 1.52 2E−07 225178_at tetratricopeptide repeat domain 14 TTC14 1.52 1E−08 204733_at “kallikrein 6 (neurosin, zyme)” KLK6 1.52 8E−07 232963_at Constitutive photomorphogenic protein COP1 1.52 1E−06 221874_at maba1 KIAA1324 1.51 2E−05 236061_at PR domain containing 15 PRDM15 1.51 2E−05 227499_at Transcribed locus NA 1.51 2E−06 241923_x_at NA NA 1.51 3E−06 208442_s_at “ataxia telangiectasia mutated (includes complementation groups ATM 1.51 3E−07 A, C and D)” 205013_s_at adenosine A2a receptor ADORA2A 1.51 1E−12 223325_at thioredoxin domain containing 11 TXNDC11 1.51 3E−10 1552586_at “transient receptor potential cation channel, subfamily V, member 3” TRPV3 1.51 5E−06 232234_at Src-like-adaptor 2 SLA2 1.51 1E−06 232706_s_at hypothetical protein PP2447 PP2447 1.51 1E−05 1557718_at “protein phosphatase 2, regulatory subunit B (B56), gamma isoform” PPP2R5C 1.51 2E−07 207798_s_at ataxin 2-like ATXN2L 1.51 1E−05 221680_s_at ets variant gene 7 (TEL2 oncogene) ETV7 1.51 4E−09 242174_at Zinc finger and BTB domain containing 10 ZBTB10 1.51 3E−05 1569490_at fibronectin type III domain containing 3B FNDC3B 1.51 2E−07 1555407_s_at “FGD1 family, member 3” FGD3 1.51 6E−08 1555734_x_at “adaptor-related protein complex 1, sigma 3 subunit” AP1S3 1.51 3E−05 243570_at KIAA0102 gene product KIAA0102 1.51 7E−09 1553067_a_at gonadotropin-releasing hormone (type 2) receptor 2 GNRHR2 1.51 2E−05 211079_s_at dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 1A | DYRK1A 1.51 1E−06 dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 1A 230551_at “Shinc-4 mRNA, partial sequence” NA 1.51 3E−05 1552554_a_at “caspase recruitment domain family, member 12” CARD12 1.51 1E−05 212079_s_at “myeloid/lymphoid or mixed-lineage leukemia MLL 1.51 3E−09 (trithorax homolog, Drosophila)” 205563_at KiSS-1 metastasis-suppressor KISS1 1.51 4E−09 217823_s_at “ubiquitin-conjugating enzyme E2, J1 (UBC6 homolog, yeast)” UBE2J1 1.51 7E−11 218796_at chromosome 20 open reading frame 42 C20orf42 1.50 4E−05 233403_x_at transmembrane 6 superfamily member 2 TM6SF2 1.50 2E−06 221297_at “G protein-coupled receptor, family C, group 5, member D” GPRC5D 1.50 2E−06 231210_at hypothetical protein LOC283129 LOC283129 1.50 6E−05 211367_s_at “caspase 1, apoptosis-related cysteine protease (interleukin 1, CASP1 1.50 4E−08 beta, convertase)” 224044_at “ras homolog gene family, member T1” RHOT1 1.50 2E−08 220216_at hypothetical protein FLJ11267 FLJ11267 1.50 3E−09 239162_at NA NA 1.50 5E−06 222729_at “F-box and WD-40 domain protein 7 (archipelago homolog, Drosophila)” FBXW7 1.50 2E−11 217824_at “ubiquitin-conjugating enzyme E2, J1 (UBC6 homolog, yeast)” UBE2J1 1.50 6E−08 1559500_at KIAA0804 protein KIAA0804 1.50 2E−06 1555772_a_at cell division cycle 25A CDC25A 1.50 9E−05 206216_at serine/threonine kinase 23 STK23 1.50 2E−11 224882_at acetyl-Coenzyme A synthetase 2 (AMP forming)-like ACAS2L 1.50 1E−10 218751_s_at “F-box and WD-40 domain protein 7 (archipelago homolog, Drosophila)” FBXW7 1.50 2E−12 207102_at “aldo-keto reductase family 1, member D1 AKR1D1 1.50 4E−05 (delta 4-3-ketosteroid-5-beta-reductase)” 226132_s_at hypothetical protein FLJ31434 FLJ31434 1.50 3E−08 213297_at hypothetical protein FLJ22318 FLJ22318 1.50 6E−12 208486_at dopamine receptor D5 DRD5 1.50 9E−06

TABLE 6 Gene expression associated with the M subtype of RA. NETAFFX: Fold ProbeID NETAFFX: GeneTitle GeneSymbol Change p-value 231755_at “interleukin 1 family, member 8 (eta)” IL1F8 15.91 9E−05 206239_s_at “serine protease inhibitor, Kazal type 1” SPINK1 10.07 4E−04 206067_s_at Wilms tumor 1 WT1 9.44 5E−08 204580_at matrix metalloproteinase 12 (macrophage elastase) MMP12 8.30 5E−07 1552394_a_at hypothetical protein FLJ25421 FLJ25421 7.89 2E−04 214974_x_at chemokine (C-X-C motif) ligand 5 CXCL5 7.83 8E−10 219890_at “C-type lectin domain family 5, member A” CLEC5A 7.36 2E−07 215101_s_at chemokine (C-X-C motif) ligand 5 CXCL5 7.34 2E−07 212657_s_at interleukin 1 receptor antagonist IL1RN 6.79 4E−08 1552393_at hypothetical protein FLJ25421 FLJ25421 6.11 6E−04 230966_at interleukin 4 induced 1 IL4I1 4.82 5E−09 216243_s_at interleukin 1 receptor antagonist IL1RN 4.65 2E−08 209696_at “fructose-1,6-bisphosphatase 1” FBP1 4.30 3E−08 211506_s_at interleukin 8 IL8 4.22 1E−06 206881_s_at “leukocyte immunoglobulin-like receptor, LILRA3 3.95 6E−06 subfamily A (without TM domain), member 3” 232629_at prokineticin 2 PROK2 3.81 2E−04 219874_at “solute carrier family 12 (potassium/chloride SLC12A8 3.77 1E−09 transporters), member 8” 212659_s_at interleukin 1 receptor antagonist IL1RN 3.72 3E−07 205220_at G protein-coupled receptor 109B | G protein- GPR109B 3.71 4E−05 coupled receptor 109B 210842_at neuropilin 2 NRP2 3.64 2E−05 216015_s_at cold autoinflammatory syndrome 1 CIAS1 3.61 2E−09 230147_at coagulation factor II (thrombin) receptor-like 2 F2RL2 3.45 8E−07 217078_s_at CD300A antigen CD300A 3.30 4E−07 202628_s_at “serine (or cysteine) proteinase inhibitor, clade E (nexin, SERPINE1 3.27 1E−07 plasminogen activator inhibitor type 1), member 1” 216953_s_at Wilms tumor 1 WT1 3.26 5E−04 202859_x_at interleukin 8 IL8 3.19 2E−05 211527_x_at vascular endothelial growth factor VEGF 3.03 2E−13 223484_at normal mucosa of esophagus specific 1 NMES1 3.03 7E−07 216598_s_at chemokine (C-C motif) ligand 2 CCL2 2.99 3E−13 203290_at “major histocompatibility complex, class II, DQ alpha 1 | HLA-DQA1 | 2.94 3E−04 major histocompatibility complex, class II, DQ alpha 1 | HLA-DQA2 major histocompatibility complex, class II, DQ alpha 2 | major histocompatibility complex, class II, DQ alpha 2” 208606_s_at “wingless-type MMTV integration site family, member 4 | WNT4 2.93 5E−04 wingless-type MMTV integration site family, member 4” 214014_at CDC42 effector protein (Rho GTPase binding) 2 CDC42EP2 2.90 4E−04 209122_at adipose differentiation-related protein ADFP 2.90 7E−06 223767_at G protein-coupled receptor 84 GPR84 2.84 9E−08 202219_at “solute carrier family 6 (neurotransmitter transporter, creatine), SLC6A8 2.77 5E−05 member 8” 211924_s_at “plasminogen activator, urokinase receptor | plasminogen PLAUR 2.74 7E−12 activator, urokinase receptor” 205709_s_at CDP-diacylglycerol synthase (phosphatidate CDS1 2.69 7E−07 cytidylyltransferase) 1 209949_at “neutrophil cytosolic factor 2 (65 kDa, chronic granulomatous NCF2 2.68 1E−09 disease, autosomal 2)” 230030_at heparan sulfate 6-O-sulfotransferase 2 HS6ST2 2.66 6E−04 213831_at “major histocompatibility complex, class II, DQ alpha 1” HLA-DQA1 2.65 1E−04 213338_at Ras-induced senescence 1 RIS1 2.65 4E−12 210845_s_at “plasminogen activator, urokinase receptor” PLAUR 2.60 8E−13 218498_s_at ERO1-like (S. cerevisiae) ERO1L 2.60 6E−09 220023_at apolipoprotein B48 receptor APOB48R 2.59 6E−08 210512_s_at vascular endothelial growth factor VEGF 2.56 5E−11 205114_s_at “chemokine (C-C motif) ligand 3 | chemokine (C-C motif) CCL3 | 2.53 2E−04 ligand 3-like 1 | chemokine (C-C motif) ligand 3-like, CCL3L1 | centromeric” MGC12815 214038_at chemokine (C-C motif) ligand 8 CCL8 2.53 6E−07 206482_at PTK6 protein tyrosine kinase 6 PTK6 2.50 4E−06 224762_at tumor differentially expressed 2-like TDE2L 2.46 4E−06 219386_s_at SLAM family member 8 SLAMF8 2.45 5E−08 205648_at wingless-type MMTV integration site family member 2 WNT2 2.43 7E−05 204533_at chemokine (C-X-C motif) ligand 10 CXCL10 2.40 4E−07 201313_at “enolase 2 (gamma, neuronal)” ENO2 2.39 5E−06 202627_s_at “serine (or cysteine) proteinase inhibitor, clade E (nexin, SERPINE1 2.37 7E−07 plasminogen activator inhibitor type 1), member 1” 204656_at SHB (Src homology 2 domain containing) adaptor protein B SHB 2.35 4E−10 214435_x_at v-ral simian leukemia viral oncogene homolog A (ras related) RALA 2.34 9E−08 212171_x_at vascular endothelial growth factor VEGF 2.32 2E−14 220016_at hypothetical protein MGC5395 MGC5395 2.30 4E−06 202638_s_at “intercellular adhesion molecule 1 (CD54), human ICAM1 2.30 2E−17 rhinovirus receptor” 217159_x_at sialic acid binding Ig-like lectin 7 SIGLEC7 2.28 5E−04 201952_at NA NA 2.25 3E−09 1555214_a_at “C-type lectin domain family 7, member A” CLEC7A 2.25 2E−04 229309_at “Adrenergic, beta-1-, receptor” ADRB1 2.24 2E−08 222646_s_at ERO1-like (S. cerevisiae) ERO1L 2.24 2E−07 212374_at fem-1 homolog b (C. elegans) FEM1B 2.23 2E−05 202856_s_at “solute carrier family 16 (monocarboxylic acid SLC16A3 2.23 4E−10 transporters), member 3” 205071_x_at X-ray repair complementing defective repair in XRCC4 2.23 6E−05 Chinese hamster cells 4 219047_s_at hypothetical protein FLJ13479 FLJ13479 2.22 6E−06 213324_at v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene SRC 2.22 2E−08 homolog (avian) 206503_x_at promyelocytic leukemia PML 2.20 2E−05 242871_at membrane progestin receptor gamma MPRG 2.20 1E−04 208438_s_at Gardner-Rasheed feline sarcoma viral (v-fgr) FGR 2.19 2E−09 oncogene homolog 205875_s_at three prime repair exonuclease 1 TREX1 2.18 5E−04 220066_at “caspase recruitment domain family, member 15” CARD15 2.18 3E−08 201951_at Activated leukocyte cell adhesion molecule ALCAM 2.17 8E−10 210233_at interleukin 1 receptor accessory protein IL1RAP 2.14 8E−10 202637_s_at “intercellular adhesion molecule 1 (CD54), human ICAM1 2.13 8E−14 rhinovirus receptor” 217818_s_at “actin related protein 2/3 complex, subunit 4, 20 kDa” ARPC4 2.12 6E−04 211013_x_at promyelocytic leukemia PML 2.12 1E−05 209933_s_at CD3OOA antigen CD300A 2.12 1E−06 220333_at membrane progestin receptor gamma MPRG 2.12 4E−06 205786_s_at “integrin, alpha M (complement component receptor 3, alpha; ITGAM 2.10 2E−09 also known as CD11b (p170), macrophage antigen alpha polypeptide) | integrin, alpha M (complement component receptor 3, alpha; also known as CD11b (p170), macrophage antigen alpha polypeptide . . .” 204174_at arachidonate 5-lipoxygenase-activating protein ALOX5AP 2.10 1E−06 205722_s_at GDNF family receptor alpha 2 GFRA2 2.10 6E−04 209875_s_at “secreted phosphoprotein 1 (osteopontin, bone sialoprotein I, SPP1 2.09 3E−05 early T-lymphocyte activation 1)” 223303_at UNC-112 related protein 2 URP2 2.09 4E−08 226111_s_at zinc finger protein 385 ZNF385 2.09 2E−09 223019_at chromosome 9 open reading frame 88 C9orf88 2.08 4E−12 215485_s_at “intercellular adhesion molecule 1 (CD54), human rhinovirus ICAM1 2.07 1E−12 receptor” 219434_at triggering receptor expressed on myeloid cells 1 TREM1 2.05 3E−08 214841_at cornichon homolog 3 (Drosophila) CNIH3 2.05 7E−06 221698_s_at “C-type lectin domain family 7, member A | C-type lectin CLEC7A 2.05 5E−09 domain family 7, member A” 223855_s_at G protein-coupled receptor 24 GPR24 2.04 1E−04 202790_at claudin 7 CLDN7 2.04 4E−06 1554952_s_at “NACHT, leucine rich repeat and PYD containing 12” NALP12 2.03 9E−09 205241_at SCO cytochrome oxidase deficient homolog 2 (yeast) SCO2 2.03 1E−13 207387_s_at glycerol kinase GK 2.02 4E−08 201250_s_at “solute carrier family 2 (facilitated glucose transporter), SLC2A1 2.02 1E−05 member 1” 206267_s_at megakaryocyte-associated tyrosine kinase MATK 2.02 1E−05 238423_at synaptotagmin-like 3 SYTL3 2.01 2E−05 1555349_a_at “integrin, beta 2 (antigen CD18 (p95), lymphocyte function- ITGB2 2.01 2E−06 associated antigen 1; macrophage antigen 1 (mac-1) beta subunit)” 221908_at Hypothetical protein FLJ14627 FLJ14627 2.00 6E−05 243611_at hypothetical protein FLJ14966 FLJ14966 1.99 4E−05 221779_at MICAL-like 1 MICAL-L1 1.99 4E−07 209053_s_at Wolf-Hirschhorn syndrome candidate 1 WHSC1 1.98 5E−04 202855_s_at “solute carrier family 16 (monocarboxylic acid transporters), SLC16A3 1.98 3E−08 member 3” 226088_at “zinc finger, DHHC domain containing 12” ZDHHC12 1.98 8E−06 1555167_s_at pre-B-cell colony enhancing factor 1 PBEF1 1.98 2E−04 218223_s_at CK2 interacting protein 1; HQ0024c protein CKIP-1 1.97 9E−10 204508_s_at carbonic anhydrase XII CA12 1.96 2E−06 205721_at GDNF family receptor alpha 2 GFRA2 1.96 4E−06 235568_at hypothetical protein LOC199675 LOC199675 1.95 9E−06 211312_s_at WNT1 inducible signaling pathway protein 1 WISP1 1.95 7E−04 205098_at chemokine (C-C motif) receptor 1 CCR1 1.95 6E−11 237623_at Cystatin C (amyloid angiopathy and cerebral hemorrhage) CST3 1.95 4E−05 218567_x_at dipeptidylpeptidase 3 DPP3 1.95 3E−04 214866_at “plasminogen activator, urokinase receptor” PLAUR 1.94 6E−11 204446_s_at arachidonate 5-lipoxygenase ALOX5 1.94 2E−06 209660_at “transthyretin (prealbumin, amyloidosis type I)” TTR 1.93 8E−06 214366_s_at arachidonate 5-lipoxygenase ALOX5 1.92 1E−05 205485_at ryanodine receptor 1 (skeletal) RYR1 1.92 1E−06 203963_at carbonic anhydrase XII CA12 1.92 6E−07 202912_at adrenomedullin ADM 1.91 5E−04 55081_at MICAL-like 1 MICAL-L1 1.91 1E−09 214164_x_at carbonic anhydrase XII CA12 1.91 1E−08 219978_s_at nucleolar and spindle associated protein 1 NUSAP1 1.90 6E−04 202910_s_at CD97 antigen CD97 1.90 5E−09 232608_x_at “caspase recruitment domain family, member 14” CARD14 1.90 3E−04 210152_at “leukocyte immunoglobulin-like receptor, subfamily B LILRB4 1.90 2E−06 (with TM and ITIM domains), member 4” 1555756_a_at “C-type lectin domain family 7, member A” CLEC7A 1.89 1E−06 225316_at hypothetical protein FLJ14490 FLJ14490 1.89 7E−04 219634_at carbohydrate (chondroitin 4) sulfotransferase 11 CHST11 1.89 1E−09 205099_s_at chemokine (C-C motif) receptor 1 CCR1 1.88 4E−08 226869_at Full length insert cDNA clone ZD77F06 NA 1.88 5E−08 1553133_at chromosome 9 open reading frame 72 C9orf72 1.88 7E−07 228302_x_at calcium/calmodulin-dependent protein kinase II CaMKIINalpha 1.88 3E−04 205165_at “cadherin, EGF LAG seven-pass G-type receptor 3 CELSR3 1.88 5E−04 (flamingo homolog, Drosophila)” 219385_at SLAM family member 8 SLAMF8 1.87 1E−05 224374_s_at elastin microfibril interfacer 2 | elastin microfibril interfacer 2 EMILIN2 1.87 1E−14 202803_s_at “integrin, beta 2 (antigen CD18 (p95), lymphocyte ITGB2 1.87 9E−07 function-associated antigen 1; macrophage antigen 1 (mac-1) beta subunit)” 207091_at “purinergic receptor P2X, ligand-gated ion channel, 7” P2RX7 1.86 1E−06 215966_x_at NA NA 1.86 3E−08 229404_at twist homolog 2 (Drosophila) TWIST2 1.86 2E−05 1559399_s_at “zinc finger, CCHC domain containing 10” ZCCHC10 1.85 4E−10 204655_at NA NA 1.85 2E−05 204490_s_at CD44 antigen (homing function and Indian blood group system) CD44 1.84 2E−10 226389_s_at Rap guanine nucleotide exchange factor (GEF) 1 RAPGEF1 1.83 5E−12 1554036_at zinc finger and BTB domain containing 24 ZBTB24 1.82 2E−04 218627_at hypothetical protein FLJ11259 FLJ11259 1.81 3E−14 214501_s_at “H2A histone family, member Y” H2AFY 1.81 2E−09 239217_x_at “ATP-binding cassette, sub-family C (CFTR/MRP), member 3” ABCC3 1.81 4E−04 217507_at “solute carrier family 11 (proton-coupled divalent metal SLC11A1 1.81 4E−05 ion transporters), member 1” 238044_at “Transcribed locus, moderately similar to XP_529341.1 NA 1.81 9E−06 LOC450700 [Pan troglodytes]” 202998_s_at lysyl oxidase-like 2 LOXL2 1.81 1E−04 202998_s_at lysyl oxidase-like 2 LOXL2 1.81 1E−04 205349_at “guanine nucleotide binding protein (G protein), GNA15 1.81 7E−09 alpha 15 (Gq class)” 206420_at “immunoglobulin superfamily, member 6” IGSF6 1.81 1E−07 226354_at “lactamase, beta” LACTB 1.81 2E−13 206991_s_at chemokine (C-C motif) receptor 5 CCR5 1.81 1E−05 215977_x_at glycerol kinase GK 1.80 4E−06 209835_x_at CD44 antigen (homing function and Indian blood CD44 1.80 2E−10 group system) 219360_s_at “transient receptor potential cation channel, subfamily TRPM4 1.80 3E−04 M, member 4” 207075_at cold autoinflammatory syndrome 1 CIAS1 1.79 1E−06 207376_at VENT-like homeobox 2 VENTX2 1.79 8E−08 200629_at tryptophanyl-tRNA synthetase WARS 1.79 6E−06 215223_s_at “superoxide dismutase 2, mitochondrial” SOD2 1.78 3E−05 214978_s_at “protein tyrosine phosphatase, receptor type, f polypeptide PPFIA4 1.78 6E−05 (PTPRF), interacting protein (liprin), alpha 4” 212014_x_at CD44 antigen (homing function and Indian blood group system) CD44 1.78 4E−09 213520_at NA NA 1.78 5E−04 209267_s_at “solute carrier family 39 (zinc transporter), member 8” SLC39A8 1.78 1E−05 223952_x_at dehydrogenase/reductase (SDR family) member 9 DHRS9 1.78 7E−05 224009_x_at dehydrogenase/reductase (SDR family) member 9 DHRS9 1.77 9E−05 215498_s_at mitogen-activated protein kinase kinase 3 | MAP2K3 1.77 8E−09 mitogen-activated protein kinase kinase 3 1554131_at KIAA1128 KIAA1128 1.76 1E−04 201890_at ribonucleotide reductase M2 polypeptide RRM2 1.76 1E−04 204440_at “CD83 antigen (activated B lymphocytes, CD83 1.74 2E−05 immunoglobulin superfamily)” 229763_at Forkhead box P4 | Activating signal cointegrator 1 FOXP4 | 1.74 3E−04 complex subunit 2 ASCC2 208018_s_at hemopoietic cell kinase HCK 1.74 2E−09 215867_x_at carbonic anhydrase XII CA12 1.74 1E−07 222817_at “hydroxy-delta-5-steroid dehydrogenase, 3 beta- HSD3B7 1.74 4E−05 and steroid delta-isomerase 7” 230283_at neuralized-like 2 (Drosophila) NEURL2 1.73 1E−04 203570_at lysyl oxidase-like 1 LOXL1 1.73 1E−07 211725_s_at BH3 interacting domain death agonist | BID 1.73 1E−12 BH3 interacting domain death agonist 223398_at chromosome 9 open reading frame 89 C9orf89 1.73 3E−09 209193_at NA NA 1.73 1E−07 205640_at “aldehyde dehydrogenase 3 family, member B1” ALDH3B1 1.73 2E−05 229975_at Transcribed locus NA 1.73 5E−04 1552914_a_at B7 homolog 3 B7H3 1.72 4E−04 207697_x_at “leukocyte immunoglobulin-like receptor, subfamily B LILRB2 1.72 2E−07 (with TM and ITIM domains), member 2” 210146_x_at “leukocyte immunoglobulin-like receptor, subfamily B LILRB2 | 1.71 7E−06 (with TM and ITIM domains), member 2 | leukocyte LILRB6 immunoglobulin-like receptor, subfamily B (with TM and ITIM domains), member 6” 212268_at “serine (or cysteine) proteinase inhibitor, clade B SERPINB1 1.71 3E−14 (ovalbumin), member 1” 219799_s_at dehydrogenase/reductase (SDR family) member 9 DHRS9 1.71 2E−04 213693_s_at “Mucin 1, transmembrane” MUC1 1.70 2E−04 202207_at ADP-ribosylation factor-like 7 ARL7 1.70 7E−08 210042_s_at cathepsin Z CTSZ 1.70 1E−06 1557458_s_at SHB (Src homology 2 domain containing) adaptor protein B SHB 1.70 2E−10 204182_s_at zinc finger protein 297B ZNF297B 1.70 2E−06 214500_at “H2A histone family, member Y” H2AFY 1.69 2E−10 209191_at “tubulin, beta 6” TUBB6 1.69 4E−09 218169_at Vac14 homolog (S. cerevisiae) VAC14 1.69 2E−07 204401_at “potassium intermediate/small conductance KCNN4 1.69 2E−06 calcium-activated channel, subfamily N, member 4” 226722_at “family with sequence similarity 20, member C” FAM20C 1.69 9E−06 217739_s_at pre-B-cell colony enhancing factor 1 PBEF1 1.69 1E−04 225440_at 1-acylglycerol-3-phosphate O-acyltransferase 3 AGPAT3 1.68 7E−06 203835_at glycoprotein A repetitions predominant GARP 1.68 2E−05 217388_s_at kynureninase (L-kynurenine hydrolase) KYNU 1.68 2E−05 219631_at low density lipoprotein-related protein 12 LRP12 1.68 1E−08 213942_at “EGF-like-domain, multiple 3” EGFL3 1.68 3E−05 212472_at flavoprotein oxidoreductase MICAL2 MICAL2 1.67 3E−04 212472_at flavoprotein oxidoreductase MICAL2 MICAL2 1.67 3E−04 225687_at chromosome 20 open reading frame 129 C20orf129 1.67 2E−04 223502_s_at “tumor necrosis factor (ligand) superfamily, member 13b” TNFSF13B 1.66 9E−06 213274_s_at cathepsin B CTSB 1.66 2E−09 228499_at “6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 4” PFKFB4 1.66 9E−05 224880_at v-ral simian leukemia viral oncogene homolog A (ras related) RALA 1.66 2E−09 209230_s_at p8 protein (candidate of metastasis 1) P8 1.66 6E−04 204879_at lung type-I cell membrane-associated glycoprotein T1A-2 1.65 3E−05 219763_at KIAA1608 KIAA1608 1.65 1E−08 201389_at “integrin, alpha 5 (fibronectin receptor, alpha polypeptide)” ITGA5 1.65 2E−06 206342_x_at iduronate 2-sulfatase (Hunter syndrome) IDS 1.65 7E−09 218840_s_at NAD synthetase 1 NADSYN1 1.64 4E−04 224282_s_at 1-acylglycerol-3-phosphate O-acyltransferase 3 AGPAT3 1.64 6E−05 228124_at chromosome 20 open reading frame 22 C20orf22 1.64 5E−07 204445_s_at arachidonate 5-lipoxygenase ALOX5 1.64 5E−04 212488_at “Collagen, type V, alpha 1” COL5A1 1.64 8E−04 212488_at “Collagen, type V, alpha 1” COL5A1 1.64 8E−04 210889_s_at “Fc fragment of IgG, low affinity IIb, receptor (CD32)” FCGR2B 1.64 2E−04 202788_at mitogen-activated protein kinase-activated protein kinase 3 MAPKAPK3 1.64 2E−09 210513_s_at vascular endothelial growth factor VEGF 1.64 2E−09 217167_x_at glycerol kinase GK 1.63 4E−05 202626_s_at v-yes-1 Yamaguchi sarcoma viral related oncogene homolog | LYN 1.63 1E−08 v-yes-1 Yamaguchi sarcoma viral related oncogene homolog 207765_s_at KIAA1539 KIAA1539 1.63 7E−09 203671_at thiopurine S-methyltransferase TPMT 1.63 2E−11 1557915_s_at glutathione S-transferase omega 1 GSTO1 1.63 1E−10 219448_at hypothetical protein FLJ20533 FLJ20533 1.63 5E−06 204158_s_at “T-cell, immune regulator 1, ATPase, H+ transporting, TCIRG1 1.63 5E−06 lysosomal V0 protein a isoform 3” 210735_s_at carbonic anhydrase XII CA12 1.63 4E−06 207939_x_at “RNA binding protein S1, serine-rich domain” RNPS1 1.63 1E−06 201231_s_at “enolase 1, (alpha)” ENO1 1.63 2E−11 221882_s_at transmembrane protein 8 (five membrane-spanning domains) TMEM8 1.63 2E−07 219053_s_at Hypothetical protein FLJ20847 FLJ20847 1.62 4E−14 219648_at likely ortholog of mouse dilute suppressor DSU 1.62 9E−05 204924_at toll-like receptor 2 TLR2 1.62 4E−05 223683_at “zinc finger, MYND domain containing 15” ZMYND15 1.61 8E−06 218309_at calcium/calmodulin-dependent protein kinase II CaMKIINalpha 1.61 1E−04 205205_at “v-rel reticuloendotheliosis viral oncogene homolog B, RELB 1.61 4E−11 nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 (avian)” 1557905_s_at CD44 antigen (homing function and Indian blood CD44 1.61 1E−06 group system) 213113_s_at “solute carrier family 43, member 3” SLC43A3 1.61 2E−10 218779_x_at EPS8-like 1 EPS8L1 1.61 4E−04 205174_s_at glutaminyl-peptide cyclotransferase (glutaminyl cyclase) QPCT 1.60 3E−04 227396_at “Homo sapiens, clone IMAGE: 4454331, mRNA” NA 1.60 7E−06 203561_at “Fc fragment of IgG, low affinity IIa, receptor (CD32)” FCGR2A 1.60 1E−05 212849_at axin 1 AXIN1 1.60 5E−08 229055_at Transcribed locus NA 1.60 6E−08 211057_at receptor tyrosine kinase-like orphan receptor 1 | ROR1 1.60 1E−04 receptor tyrosine kinase-like orphan receptor 1 226944_at HtrA serine peptidase 3 HTRA3 1.60 3E−05 202206_at ADP-ribosylation factor-like 7 ARL7 1.59 2E−06 201743_at CD14 antigen | CD14 antigen CD14 1.59 3E−08 230498_at G protein-coupled receptor 24 GPR24 1.59 5E−07 210612_s_at synaptojanin 2 SYNJ2 1.59 6E−04 210612_s_at synaptojanin 2 SYNJ2 1.59 6E−04 35820_at GM2 ganglioside activator GM2A 1.58 3E−07 1553375_at BTB (POZ) domain containing 9 BTBD9 1.58 7E−04 221042_s_at “calmin (calponin-like, transmembrane)” CLMN 1.58 1E−05 217995_at sulfide quinone reductase-like (yeast) SQRDL 1.58 9E−07 1554406_a_at “C-type lectin domain family 7, member A” CLEC7A 1.58 1E−07 235735_at NA NA 1.58 6E−06 1560060_s_at vacuolar protein sorting 37C (yeast) VPS37C 1.58 4E−13 207540_s_at spleen tyrosine kinase SYK 1.58 6E−05 218145_at tribbles homolog 3 (Drosophila) TRIB3 1.58 1E−04 204657_s_at SHB (Src homology 2 domain containing) adaptor protein B SHB 1.58 7E−07 224950_at prostaglandin F2 receptor negative regulator PTGFRN 1.57 1E−05 232510_s_at dipeptidylpeptidase 3 DPP3 1.57 2E−07 1554503_a_at osteoclast-associated receptor OSCAR 1.57 1E−04 203952_at activating transcription factor 6 ATF6 1.57 1E−04 226497_s_at Fms-related tyrosine kinase 1 (vascular endothelial growth FLT1 1.57 5E−05 factor/vascular permeability factor receptor) 201470_at glutathione S-transferase omega 1 GSTO1 1.56 2E−10 219090_at “solute carrier family 24 (sodium/potassium/calcium SLC24A3 1.56 1E−03 exchanger), member 3” 205707_at interleukin 17 receptor IL17R 1.56 2E−06 220253_s_at low density lipoprotein-related protein 12 LRP12 1.56 8E−07 213011_s_at triosephosphate isomerase 1 TPI1 1.56 8E−10 227060_at “tumor necrosis factor receptor superfamily, member 19-like” TNFRSF19L 1.56 9E−04 207671_s_at “vitelliform macular dystrophy (Best disease, bestrophin)” VMD2 1.56 3E−04 227757_at cullin 4A CUL4A 1.56 3E−04 226140_s_at OTU domain containing 1 OTUD1 1.56 8E−10 209545_s_at receptor-interacting serine-threonine kinase 2 RIPK2 1.56 3E−09 218009_s_at protein regulator of cytokinesis 1 PRC1 1.56 1E−04 202208_s_at ADP-ribosylation factor-like 7 ARL7 1.56 1E−06 203045_at ninjurin 1 NINJ1 1.56 4E−07 207168_s_at “H2A histone family, member Y” H2AFY 1.55 9E−10 221654_s_at ubiquitin specific protease 3 USP3 1.55 2E−09 203402_at “potassium voltage-gated channel, shaker-related subfamily, KCNAB2 1.55 3E−05 beta member 2” 203923_s_at “cytochrome b-245, beta polypeptide (chronic CYBB 1.55 1E−04 granulomatous disease)” 202897_at “protein tyrosine phosphatase, non-receptor type substrate 1” PTPNS1 1.55 3E−09 201848_s_at BCL2/adenovirus E1B 19 kDa interacting protein 3 BNIP3 1.55 3E−06 202625_at V-yes-1 Yamaguchi sarcoma viral related oncogene homolog | LYN 1.55 2E−09 V-yes-1 Yamaguchi sarcoma viral related oncogene homolog 1565951_s_at choroideremia-like (Rab escort protein 2) CHML 1.55 1E−04 41660_at “cadherin, EGF LAG seven-pass G-type receptor 1 CELSR1 1.55 7E−04 (flamingo homolog, Drosophila)” 215499_at Mitogen-activated protein kinase kinase 3 | MAP2K3 1.55 2E−07 Mitogen-activated protein kinase kinase 3 203409_at “damage-specific DNA binding protein 2, 48 kDa” DDB2 1.55 2E−06 224733_at chemokine-like factor super family 3 CKLFSF3 1.55 4E−10 204858_s_at endothelial cell growth factor 1 (platelet-derived) ECGF1 1.54 4E−05 201849_at BCL2/adenovirus E1B 19 kDa interacting protein 3 BNIP3 1.54 3E−06 225955_at “meteorin, glial cell differentiation regulator-like” METRNL 1.54 2E−08 222768_s_at CGI-09 protein CGI-09 1.54 3E−09 204575_s_at matrix metalloproteinase 19 MMP19 1.54 2E−05 208002_s_at brain acyl-CoA hydrolase BACH 1.54 2E−06 225738_at Rap guanine nucleotide exchange factor (GEF) 1 RAPGEF1 1.54 2E−11 1553134_s_at chromosome 9 open reading frame 72 C9orf72 1.54 4E−04 227337_at low density lipoprotein receptor-related protein binding protein Lrp2bp 1.53 2E−04 221539_at eukaryotic translation initiation factor 4E binding protein 1 EIF4EBP1 1.53 9E−05 216180_s_at synaptojanin 2 SYNJ2 1.53 4E−04 235678_at GM2 ganglioside activator GM2A 1.53 2E−07 202087_s_at cathepsin L CTSL 1.53 2E−06 213716_s_at secreted and transmembrane 1 SECTM1 1.53 2E−05 201963_at acyl-CoA synthetase long-chain family member 1 ACSL1 1.53 5E−06 223811_s_at chromosome 7 open reading frame 20 C7orf20 1.53 7E−05 212671_s_at “major histocompatibility complex, class II, DQ alpha 1 | HLA-DQA1 | major histocompatibility complex, class II, DQ alpha 2” HLA-DQA2 1.53 9E−04 207275_s_at acyl-CoA synthetase long-chain family member 1 ACSL1 1.53 3E−04 209166_s_at “mannosidase, alpha, class 2B, member 1” MAN2B1 1.53 6E−08 208308_s_at glucose phosphate isomerase GPI 1.53 1E−07 202329_at c-src tyrosine kinase CSK 1.53 1E−06 205684_s_at chromosome 9 open reading frame 55 C9orf55 1.53 1E−08 210589_s_at “glucosidase, beta; acid (includes glucosylceramidase)” GBA 1.52 9E−05 222868_s_at interleukin 18 binding protein IL18BP 1.52 6E−06 201587_s_at interleukin-1 receptor-associated kinase 1 IRAK1 1.52 3E−09 207224_s_at sialic acid binding Ig-like lectin 7 SIGLEC7 1.52 1E−04 1554309_at “eukaryotic translation initiation factor 4 gamma, 3” EIF4G3 1.52 4E−04 228153_at IBR domain containing 2 IBRDC2 1.52 7E−07 217984_at ribonuclease T2 RNASET2 1.52 4E−05 200840_at lysyl-tRNA synthetase KARS 1.52 4E−07 218039_at nucleolar and spindle associated protein 1 NUSAP1 1.52 1E−04 226459_at phosphoinositide-3-kinase adaptor protein 1 PIK3AP1 1.52 1E−06 224937_at prostaglandin F2 receptor negative regulator PTGFRN 1.51 3E−04 224937_at prostaglandin F2 receptor negative regulator PTGFRN 1.51 3E−04 201079_at synaptogyrin 2 SYNGR2 1.51 1E−05 242288_s_at elastin microfibril interfacer 2 EMILIN2 1.51 1E−05 201761_at “methylenetetrahydrofolate dehydrogenase (NADP+ MTHFD2 1.51 2E−08 dependent) 2, methenyltetrahydrofolate cyclohydrolase” 226071_at thrombospondin repeat containing 1 TSRC1 1.51 5E−07 224859_at B7 homolog 3 B7H3 1.51 1E−05 214175_x_at PDZ and LIM domain 4 PDLIM4 1.51 1E−03 223378_at NA NA 1.51 2E−05 202009_at PTK9L protein tyrosine kinase 9-like (A6-related protein) PTK9L 1.51 3E−12 219040_at coronin 7 CORO7 1.51 2E−04 228745_at Hypothetical protein FLJ13611 FLJ13611 1.51 6E−08 221894_at aarF domain containing kinase 2 ADCK2 1.51 2E−05 235359_at ELLP3030 UNQ3030 1.50 7E−05 217933_s_at leucine aminopeptidase 3 LAP3 1.50 1E−05 1555841_at similar to RIKEN cDNA 5730528L13 gene MGC17337 1.50 9E−08 201779_s_at ring finger protein 13 RNF13 1.50 1E−08 201066_at cytochrome c-1 CYC1 1.50 4E−08

TABLE 7 Gene expression associated with the F2 subtype of RA. NETAFFX: Fold ProbeID NETAFFX: GeneTitle GeneSymbol Change p-value 206552_s_at “tachykinin, precursor 1 (substance K, substance P, TAC1 10.84 3E−05 neurokinin 1, neurokinin 2, neuromedin L, neurokinin alpha, neuropeptide K, neuropeptide gamma)” 204712_at WNT inhibitory factor 1 WIF1 8.21 5E−07 241412_at betacellulin BTC 5.75 2E−12 223836_at Ksp37 protein KSP37 5.58 2E−08 237466_s_at hedgehog interacting protein HHIP 5.34 1E−07 206423_at angiopoietin-like 7 ANGPTL7 5.31 3E−06 206315_at cytokine receptor-like factor 1 CRLF1 4.87 5E−20 207326_at betacellulin BTC 4.48 7E−11 209613_s_at “alcohol dehydrogenase IB (class I), beta polypeptide” ADH1B 4.42 8E−05 204933_s_at “tumor necrosis factor receptor superfamily, member 11b TNFRSF11B 3.79 8E−08 (osteoprotegerin)” 214680_at “neurotrophic tyrosine kinase, receptor, type 2” NTRK2 3.78 8E−13 206262_at “alcohol dehydrogenase 1C (class I), gamma polypeptide” ADH1C 3.78 4E−06 201525_at apolipoprotein D APOD 3.74 1E−05 207174_at glypican 5 GPC5 3.71 4E−08 211276_at transcription elongation factor A (SII)-like 2 TCEAL2 3.70 4E−13 230593_at Transcribed locus NA 3.67 1E−05 231798_at noggin NOG 3.60 1E−06 223775_at hedgehog interacting protein HHIP 3.58 1E−06 206002_at G protein-coupled receptor 64 GPR64 3.52 3E−15 235050_at “solute carrier family 2 (facilitated glucose transporter), SLC2A12 3.33 1E−06 member 12” 38037_at heparin-binding EGF-like growth factor HBEGF 3.33 3E−09 221795_at “neurotrophic tyrosine kinase, receptor, type 2” NTRK2 3.29 8E−10 203662_s_at tropomodulin 1 TMOD1 3.24 1E−10 210155_at “myocilin, trabecular meshwork inducible MYOC 3.24 3E−08 glucocorticoid response” 221796_at “neurotrophic tyrosine kinase, receptor, type 2” NTRK2 3.24 1E−12 206737_at “wingless-type MMTV integration site family, member 11” WNT11 3.24 4E−07 204469_at “protein tyrosine phosphatase, receptor-type, Z polypeptide 1” PTPRZ1 3.22 8E−10 212444_at “CDNA clone IMAGE: 6025865, partial cds” NA 3.18 1E−08 213317_at Chloride intracellular channel 5 CLIC5 3.15 1E−09 1556037_s_at hedgehog interacting protein HHIP 3.14 3E−10 204932_at “tumor necrosis factor receptor superfamily, member 11b TNFRSF11B 3.09 1E−07 (osteoprotegerin)” 219295_s_at procollagen C-endopeptidase enhancer 2 PCOLCE2 3.05 5E−16 205397_x_at “SMAD, mothers against DPP homolog 3 (Drosophila)” SMAD3 2.97 2E−14 224212_s_at “protocadherin alpha 9 | protocadherin alpha subfamily C, 2 | PCDHA9 | 2.96 2E−06 protocadherin alpha subfamily C, 1 | protocadherin alpha 13 | PCDHAC2 | protocadherin alpha 12 | protocadherin alpha 11 | PCDHAC1 | protocadherin alpha 10 | protocadherin alpha 8 | PCDHA13 | protocadherin alpha 7 | prot . . .” PCDHA12 | PCDHA11 | PCDHA10 | PCDHA8 | PCDHA7 | PCDHA6 | PCDHA5 | PCDHA4 | PCDHA3 | PCDHA2 | PCDHA1 211485_s_at fibroblast growth factor 18 FGF18 2.96 1E−05 220076_at “ankylosis, progressive homolog (mouse)” ANKH 2.90 1E−10 228796_at copine IV CPNE4 2.87 5E−06 207317_s_at calsequestrin 2 (cardiac muscle) CASQ2 2.86 9E−07 214040_s_at “gelsolin (amyloidosis, Finnish type)” GSN 2.85 3E−11 206227_at “cartilage intermediate layer protein, nucleotide CILP 2.85 2E−06 pyrophosphohydrolase” 207292_s_at mitogen-activated protein kinase 7 MAPK7 2.84 3E−14 205883_at zinc finger and BTB domain containing 16 ZBTB16 2.82 3E−05 229929_at SPRY domain-containing SOCS box protein SSB-4 SSB4 2.77 1E−05 219866_at chloride intracellular channel 5 CLIC5 2.77 1E−09 213176_s_at latent transforming growth factor beta binding protein 4 LTBP4 2.75 8E−10 231729_s_at calcyphosine CAPS 2.72 8E−06 1555997_s_at insulin-like growth factor binding protein 5 IGFBP5 2.71 1E−05 227803_at ectonucleotide pyrophosphatase/phosphodiesterase 5 ENPP5 2.69 2E−08 (putative function) 206941_x_at NA NA 2.68 1E−07 227782_at Similar to B230208J24Rik protein NA 2.68 9E−18 209616_s_at carboxylesterase 1 (monocyte/macrophage serine esterase 1) CES1 2.67 5E−07 220484_at mucolipin 3 MCOLN3 2.67 6E−07 218180_s_at EPS8-like 2 EPS8L2 2.67 5E−07 1555800_at zinc finger protein 533 ZNF533 2.66 2E−06 214961_at KIAA0774 KIAA0774 2.63 4E−05 235367_at myopalladin MYPN 2.63 4E−06 230360_at collomin COLM 2.63 4E−09 229176_at “Ankylosis, progressive homolog (mouse)” ANKH 2.56 3E−10 203821_at heparin-binding EGF-like growth factor HBEGF 2.51 2E−08 217628_at Chloride intracellular channel 5 CLIC5 2.49 5E−05 203463_s_at epsin 2 EPN2 2.49 1E−05 230081_at “phosphatidylinositol-specific phospholipase C, X domain PLCXD3 2.48 2E−07 containing 3” 204442_x_at latent transforming growth factor beta binding protein 4 LTBP4 2.48 5E−09 244353_s_at NA NA 2.47 5E−07 229019_at zinc finger protein 533 ZNF533 2.47 3E−07 206404_at fibroblast growth factor 9 (glia-activating factor) FGF9 2.45 6E−06 227401_at interleukin 17D IL17D 2.45 2E−12 223093_at “ankylosis, progressive homolog (mouse)” ANKH 2.45 2E−12 204223_at proline arginine-rich end leucine-rich repeat protein PRELP 2.45 5E−09 223315_at netrin 4 NTN4 2.44 5E−15 205325_at phytanoyl-CoA hydroxylase interacting protein PHYHIP 2.42 2E−06 204714_s_at “coagulation factor V (proaccelerin, labile factor)” F5 2.42 6E−08 221926_s_at interleukin 17 receptor C IL17RC 2.41 8E−09 212741_at monoamine oxidase A MAOA 2.41 2E−08 204731_at “transforming growth factor, beta receptor III (betaglycan, TGFBR3 2.38 1E−08 300 kDa)” 219747_at hypothetical protein FLJ23191 FLJ23191 2.37 3E−14 206987_x_at fibroblast growth factor 18 FGF18 2.37 2E−05 219140_s_at “retinol binding protein 4, plasma” RBP4 2.34 3E−08 222043_at “clusterin (complement lysis inhibitor, SP-40,40, sulfated CLU 2.34 6E−13 glycoprotein 2, testosterone-repressed prostate message 2, apolipoprotein J)” 205182_s_at zinc finger protein 324 ZNF324 2.34 1E−05 211029_x_at fibroblast growth factor 18 | fibroblast growth factor 18 FGF18 2.33 3E−05 228224_at proline arginine-rich end leucine-rich repeat protein PRELP 2.33 4E−07 206614_at growth differentiation factor 5 (cartilage-derived GDF5 2.33 1E−06 morphogenetic protein-1) 1555801_s_at zinc finger protein 533 ZNF533 2.33 1E−05 1561396_at EPH receptor A6 EPHA6 2.32 3E−07 238441_at “CDNA clone IMAGE: 5288757, partial cds” NA 2.31 4E−07 228873_at “collagen, type XXII, alpha 1” COL22A1 2.30 2E−05 203068_at kelch-like 21 (Drosophila) KLHL21 2.29 2E−08 201539_s_at four and a half LIM domains 1 FHL1 2.26 1E−08 210397_at “defensin, beta 1” DEFB1 2.25 2E−05 215913_s_at “GULP, engulfment adaptor PTB domain containing 1” GULP1 2.24 6E−09 231029_at Transcribed locus NA 2.23 6E−08 203424_s_at insulin-like growth factor binding protein 5 IGFBP5 2.21 8E−05 205158_at “ribonuclease, RNase A family, 4” RNASE4 2.21 4E−16 210823_s_at “protein tyrosine phosphatase, receptor type, S” PTPRS 2.21 1E−07 204237_at “GULP, engulfment adaptor PTB domain containing 1” GULP1 2.20 3E−14 204713_s_at “coagulation factor V (proaccelerin, labile factor)” F5 2.19 7E−08 225879_at likely homolog of yeast SEN54 SEN54L 2.18 9E−09 212713_at microfibrillar-associated protein 4 MFAP4 2.18 9E−07 227762_at Transcribed locus NA 2.18 7E−05 200965_s_at actin binding LIM protein 1 ABLIM1 2.17 1E−08 224059_s_at numb homolog (Drosophila)-like NUMBL 2.16 6E−05 209355_s_at phosphatidic acid phosphatase type 2B PPAP2B 2.16 2E−09 219949_at leucine rich repeat containing 2 LRRC2 2.15 8E−10 211958_at insulin-like growth factor binding protein 5 IGFBP5 2.15 1E−07 203706_s_at frizzled homolog 7 (Drosophila) FZD7 2.15 7E−15 212062_at “ATPase, Class II, type 9A” ATP9A 2.14 1E−10 207336_at SRY (sex determining region Y)-box 5 SOX5 2.14 2E−09 203851_at insulin-like growth factor binding protein 6 IGFBP6 2.13 3E−07 205265_s_at aortic preferentially expressed protein 1 APEG1 2.13 2E−06 230083_at Ubiquitin specific protease 53 USP53 2.12 5E−10 204389_at monoamine oxidase A MAOA 2.12 5E−06 1555958_at NA NA 2.11 6E−11 205100_at glutamine-fructose-6-phosphate transaminase 2 GFPT2 2.10 1E−10 231817_at ubiquitin specific protease 53 USP53 2.10 6E−11 201801_s_at “solute carrier family 29 (nucleoside transporters), member 1” SLC29A1 2.10 9E−14 227702_at “cytochrome P450, family 4, subfamily X, polypeptide 1” CYP4X1 2.09 2E−07 206243_at tissue inhibitor of metalloproteinase 4 TIMP4 2.09 2E−05 229674_at SERTA domain containing 4 SERTAD4 2.08 8E−07 208792_s_at “clusterin (complement lysis inhibitor, SP-40,40, sulfated CLU 2.07 3E−10 glycoprotein 2, testosterone-repressed prostate message 2, apolipoprotein J)” 210298_x_at four and a half LIM domains 1 FHL1 2.07 8E−08 201149_s_at “tissue inhibitor of metalloproteinase 3 (Sorsby fundus TIMP3 2.06 1E−11 dystrophy, pseudoinflammatory)” 226192_at “Transcribed locus, strongly similar to XP_496055.1 similar NA 2.06 6E−08 to p40 [Homo sapiens]” 223094_s_at “ankylosis, progressive homolog (mouse)” ANKH 2.06 6E−07 203108_at “G protein-coupled receptor, family C, group 5, member A” GPCR5A 2.06 5E−06 203661_s_at tropomodulin 1 TMOD1 2.05 3E−11 209283_at “crystallin, alpha B” CRYAB 2.04 2E−09 224657_at mitogen-inducible gene 6 MIG-6 2.04 4E−08 224325_at frizzled homolog 8 (Drosophila) | frizzled homolog 8 FZD8 2.04 2E−13 (Drosophila) 227405_s_at frizzled homolog 8 (Drosophila) FZD8 2.03 1E−13 204736_s_at chondroitin sulfate proteoglycan 4 (melanoma-associated) CSPG4 2.03 1E−06 204235_s_at “GULP, engulfment adaptor PTB domain containing 1” GULP1 2.03 8E−12 237054_at Ectonucleotide pyrophosphatase/phosphodiesterase 5 ENPP5 2.03 3E−05 (putative function) 238489_at “Transcribed locus, moderately similar to NP_694983.1 NA 2.01 2E−05 DHHC-containing protein 20 [Homo sapiens]” 204796_at Echinoderm microtubule associated protein like 1 EML1 2.00 1E−13 220744_s_at WD repeat domain 10 WDR10 2.00 5E−05 212730_at desmuslin DMN 2.00 2E−06 204776_at thrombospondin 4 THBS4 2.00 5E−07 212328_at KIAA1102 protein KIAA1102 1.99 8E−10 220110_s_at nuclear RNA export factor 3 NXF3 1.99 1E−05 219825_at “cytochrome P450, family 26, subfamily B, polypeptide 1” CYP26B1 1.99 6E−06 229310_at kelch repeat and BTB (POZ) domain containing 9 KBTBD9 1.98 4E−16 203705_s_at frizzled homolog 7 (Drosophila) FZD7 1.98 1E−13 213800_at complement factor H CFH 1.98 1E−10 209905_at homeo box A9 HOXA9 1.96 4E−07 205591_at olfactomedin 1 OLFM1 1.96 2E−10 227821_at “leucine-rich repeat LGI family, member 4” LGI4 1.95 8E−05 239488_at Protein phosphatase 1 (formerly 2C)-like PPM1L 1.94 1E−06 237465_at hypothetical gene supported by BC062741 LOC401151 1.94 3E−06 205606_at low density lipoprotein receptor-related protein 6 LRP6 1.94 4E−05 231781_s_at leucine rich repeat containing 2 LRRC2 1.93 2E−09 205498_at growth hormone receptor GHR 1.93 1E−09 227892_at “CDNA clone IMAGE: 5288757, partial cds” NA 1.93 2E−05 203088_at fibulin 5 FBLN5 1.93 9E−07 220442_at UDP-N-acetyl-alpha-D-galactosamine: polypeptide GALNT4 1.92 1E−05 N-acetylgalactosaminyltransferase 4 (GalNAc-T4) 214505_s_at four and a half LIM domains 1 FHL1 1.92 5E−07 210674_s_at “protocadherin alpha 9 | protocadherin alpha subfamily C, 2 | PCDHA9 | 1.92 1E−05 protocadherin alpha subfamily C, 1 | protocadherin alpha 13 | PCDHAC2 | protocadherin alpha 12 | protocadherin alpha 11 | PCDHAC1 | protocadherin alpha 10 | protocadherin alpha 8 | PCDHA13 | protocadherin alpha 7 | prot . . .” PCDHA12 | PCDHA11 | PCDHA10 | PCDHA8 | PCDHA7 | PCDHA6 | PCDHA5 | PCDHA4 | PCDHA3 | PCDHA2 | PCDHA1 201148_s_at “tissue inhibitor of metalloproteinase 3 (Sorsby fundus TIMP3 1.92 4E−12 dystrophy, pseudoinflammatory)” 212327_at KIAA1102 protein KIAA1102 1.91 7E−13 204396_s_at G protein-coupled receptor kinase 5 GRK5 1.91 1E−15 218532_s_at hypothetical protein FLJ20152 FLJ20152 1.90 3E−11 231778_at distal-less homeo box 3 DLX3 1.90 5E−07 227058_at hypothetical protein FLJ14834 FLJ14834 1.90 7E−11 202686_s_at AXL receptor tyrosine kinase AXL 1.89 4E−13 1553706_at HtrA serine peptidase 4 HTRA4 1.89 3E−05 210473_s_at G protein-coupled receptor 125 GPR125 1.89 3E−10 218484_at NADH: ubiquinone oxidoreductase MLRQ subunit homolog LOC56901 1.89 2E−06 219764_at frizzled homolog 10 (Drosophila) FZD10 1.89 9E−10 212230_at phosphatidic acid phosphatase type 2B PPAP2B 1.88 2E−09 204578_at KIAA0377 gene product KIAA0377 1.88 2E−05 203786_s_at tumor protein D52-like 1 TPD52L1 1.88 1E−06 227341_at Chromosome 10 open reading frame 30 C10orf30 1.88 8E−06 210372_s_at tumor protein D52-like 1 TPD52L1 1.88 2E−06 221408_x_at protocadherin beta 12 PCDHB12 1.88 7E−07 212736_at chromosome 16 open reading frame 45 C16orf45 1.88 3E−12 213497_at ankyrin repeat and BTB (POZ) domain containing 2 ABTB2 1.87 2E−13 201147_s_at “tissue inhibitor of metalloproteinase 3 (Sorsby fundus TIMP3 1.87 1E−12 dystrophy, pseudoinflammatory)” 216017_s_at NGFI-A binding protein 2 (EGR1 binding protein 2) NAB2 1.87 2E−06 229797_at mucolipin 3 MCOLN3 1.87 1E−05 210619_s_at hyaluronoglucosaminidase 1 HYAL1 1.87 1E−09 221317_x_at protocadherin beta 6 PCDHB6 1.87 8E−07 201926_s_at “decay accelerating factor for complement (CD55, Cromer DAF 1.87 1E−08 blood group system)” 204388_s_at monoamine oxidase A MAOA 1.86 1E−06 215305_at “platelet-derived growth factor receptor, alpha polypeptide” PDGFRA 1.86 2E−05 211607_x_at “epidermal growth factor receptor (erythroblastic leukemia EGFR 1.86 2E−06 viral (v-erb-b) oncogene homolog, avian) | epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)” 229337_at ubiquitin specific protease 2 USP2 1.86 1E−05 204797_s_at echinoderm microtubule associated protein like 1 EML1 1.84 9E−11 204310_s_at natriuretic peptide receptor B/guanylate cyclase B NPR2 1.84 8E−11 (atrionatriuretic peptide receptor B) 225817_at cingulin-like 1 CGNL1 1.84 1E−08 205086_s_at hypothetical protein 384D8_6 384D8-2 1.83 4E−06 208202_s_at PHD finger protein 15 PHF15 1.83 5E−06 209683_at NA NA 1.83 7E−11 220043_s_at antigen p97 (melanoma associated) identified by MFI2 1.83 9E−05 monoclonal antibodies 133.2 and 96.5 205613_at B/K protein LOC51760 1.83 3E−05 216949_s_at polycystic kidney disease 1 (autosomal dominant) PKD1 1.82 2E−06 202289_s_at “transforming, acidic coiled-coil containing protein 2” TACC2 1.82 7E−12 205384_at FXYD domain containing ion transport regulator 1 FXYD1 1.82 2E−08 (phospholemman) 212226_s_at phosphatidic acid phosphatase type 2B PPAP2B 1.81 2E−06 223475_at LCCL domain containing cysteine-rich secretory protein 1 LCRISP1 1.81 3E−05 203425_s_at insulin-like growth factor binding protein 5 IGFBP5 1.81 8E−05 206850_at RAS-related on chromosome 22 RRP22 1.81 1E−08 218692_at hypothetical protein FLJ20366 FLJ20366 1.81 1E−07 208791_at “clusterin (complement lysis inhibitor, SP-40,40, sulfated CLU 1.80 1E−07 glycoprotein 2, testosterone-repressed prostate message 2, apolipoprotein J)” 218500_at mesenchymal stem cell protein DSCD75 LOC51337 1.80 3E−06 206480_at leukotriene C4 synthase LTC4S 1.80 6E−08 201188_s_at “inositol 1,4,5-triphosphate receptor, type 3” ITPR3 1.80 7E−08 224210_s_at “peroxisomal membrane protein 4, 24 kDa” PXMP4 1.80 3E−07 213397_x_at “ribonuclease, RNase A family, 4” RNASE4 1.80 6E−13 226901_at hypothetical protein LOC284018 LOC284018 1.79 1E−12 203355_s_at pleckstrin and Sec7 domain containing 3 PSD3 1.79 2E−06 225627_s_at KIAA1573 protein KIAA1573 1.79 2E−06 221272_s_at chromosome 1 open reading frame 21 | C1orf21 1.78 2E−05 chromosome 1 open reading frame 21 244623_at Transcribed locus NA 1.78 6E−05 205236_x_at “superoxide dismutase 3, extracellular” SOD3 1.78 9E−09 228255_at “amyotrophic lateral sclerosis 2 (juvenile) chromosome ALS2CR4 1.78 3E−08 region, candidate 4” 244130_at Transcribed locus NA 1.78 2E−05 201860_s_at “plasminogen activator, tissue” PLAT 1.77 1E−07 218613_at pleckstrin and Sec7 domain containing 3 PSD3 1.77 6E−08 232099_at protocadherin beta 16 PCDHB16 1.77 8E−07 1556427_s_at similar to hypothetical protein LOC221091 1.77 5E−10 206171_at adenosine A3 receptor ADORA3 1.76 3E−06 1557292_a_at mucolipin 3 MCOLN3 1.76 9E−05 229657_at Hypothetical gene supported by AK096885; AK098084 NA 1.76 1E−08 201802_at “solute carrier family 29 (nucleoside transporters), member 1” SLC29A1 1.76 2E−14 203426_s_at insulin-like growth factor binding protein 5 IGFBP5 1.76 4E−05 228716_at hypothetical gene supported by AK096885; AK098084 LOC401059 1.76 1E−07 217626_at “aldo-keto reductase family 1, member C1 (dihydrodiol AKR1C1 | 1.75 8E−06 dehydrogenase 1; 20-alpha (3-alpha)-hydroxysteroid AKR1C2 dehydrogenase) | aldo-keto reductase family 1, member C2 (dihydrodiol dehydrogenase 2; bile acid binding protein; 3-alpha hydroxysteroid dehydroge . . .” 201925_s_at “decay accelerating factor for complement (CD55, Cromer DAF 1.75 5E−07 blood group system)” 204066_s_at “centaurin, gamma 2” CENTG2 1.75 8E−06 1554008_at oncostatin M receptor OSMR 1.74 2E−05 221204_s_at cartilage acidic protein 1 CRTAC1 1.74 2E−06 210299_s_at four and a half LIM domains 1 FHL1 1.74 4E−07 229916_at ectonucleotide pyrophosphatase/phosphodiesterase 6 ENPP6 1.74 9E−07 202342_s_at tripartite motif-containing 2 TRIM2 1.74 1E−09 227899_at vitrin VIT 1.74 1E−08 228608_at Voltage gated channel like 1 VGCNL1 1.74 1E−06 206638_at 5-hydroxytryptamine (serotonin) receptor 2B HTR2B 1.74 9E−05 1553572_a_at cytoglobin CYGB 1.74 2E−06 226189_at “Homo sapiens, clone IMAGE: 4794726, mRNA” NA 1.73 5E−05 204041_at monoamine oxidase B MAOB 1.73 1E−06 238451_at “membrane protein, palmitoylated 7 (MAGUK p55 subfamily MPP7 1.73 1E−05 member 7)” 219902_at betaine-homocysteine methyltransferase 2 BHMT2 1.73 2E−08 1568868_at NA NA 1.73 4E−05 205398_s_at “SMAD, mothers against DPP homolog 3 (Drosophila)” SMAD3 1.73 5E−08 209894_at leptin receptor LEPR 1.72 3E−05 209019_s_at PTEN induced putative kinase 1 PINK1 1.72 4E−09 204284_at “protein phosphatase 1, regulatory (inhibitor) subunit 3C” PPP1R3C 1.72 3E−06 209243_s_at paternally expressed 3 PEG3 1.72 5E−06 228554_at MRNA; cDNA DKFZp586G0321 (from clone NA 1.72 3E−05 DKFZp586G0321) 220407_s_at “transforming growth factor, beta 2” TGFB2 1.71 8E−10 225871_at six transmembrane epithelial antigen of prostate 2 STEAP2 1.71 2E−07 206163_at mab-21-like 1 (C. elegans) MAB21L1 1.70 9E−05 202724_s_at forkhead box O1A (rhabdomyosarcoma) FOXO1A 1.70 5E−12 214721_x_at CDC42 effector protein (Rho GTPase binding) 4 CDC42EP4 1.70 2E−07 40524_at “protein tyrosine phosphatase, non-receptor type 21” PTPN21 1.70 6E−06 202669_s_at ephrin-B2 EFNB2 1.69 2E−05 221667_s_at heat shock 22 kDa protein 8 HSPB8 1.69 2E−06 232305_at 3-hydroxymethyl-3-methylglutaryl-Coenzyme A lyase-like 1 HMGCLL1 1.69 1E−05 200636_s_at “protein tyrosine phosphatase, receptor type, F” PTPRF 1.69 7E−07 202341_s_at tripartite motif-containing 2 TRIM2 1.69 2E−06 218510_x_at hypothetical protein FLJ20152 FLJ20152 1.69 8E−09 201984_s_at “epidermal growth factor receptor (erythroblastic EGFR 1.68 7E−08 leukemia viral (v-erb-b) oncogene homolog, avian)” 1554705_at hypothetical protein MGC45780 MGC45780 1.68 5E−05 211110_s_at androgen receptor (dihydrotestosterone receptor; testicular AR 1.68 5E−09 feminization; spinal and bulbar muscular atrophy; Kennedy disease) 1555950_a_at “decay accelerating factor for complement DAF 1.67 8E−08 (CD55, Cromer blood group system)” 227898_s_at hypothetical protein FLJ38705 FLJ38705 1.67 8E−06 202862_at fumarylacetoacetate hydrolase (fumarylacetoacetase) FAH 1.67 6E−11 214682_at Hypothetical protein LOC339047 LOC339047 1.67 4E−07 48031_r_at chromosome 5 open reading frame 4 C5orf4 1.67 1E−06 205019_s_at vasoactive intestinal peptide receptor 1 VIPR1 1.67 3E−06 209789_at “coronin, actin binding protein, 2B” CORO2B 1.67 9E−07 219922_s_at latent transforming growth factor beta binding protein 3 LTBP3 1.66 4E−10 37022_at proline arginine-rich end leucine-rich repeat protein PRELP 1.66 7E−06 235076_at nuclear domain 10 protein NDP52 1.65 6E−06 200637_s_at “protein tyrosine phosphatase, receptor type, F” PTPRF 1.65 2E−06 223092_at “ankylosis, progressive homolog (mouse)” ANKH 1.64 1E−07 227419_x_at placenta-specific 9 PLAC9 1.64 4E−07 236325_at NA NA 1.64 1E−06 212494_at tensin like C1 domain containing phosphatase TENC1 1.64 2E−09 226602_s_at similar to breakpoint cluster region isoform 1 LOC440820 1.64 2E−12 223796_at contactin associated protein-like 3 | similar to cell CNTNAP3 | 1.63 5E−06 recognition molecule CASPR3 LOC389734 229487_at Early B-cell factor EBF 1.63 4E−07 225867_at “mental retardation, X-linked 85” MRX85 1.63 2E−08 204036_at “endothelial differentiation, lysophosphatidic acid EDG2 1.63 6E−10 G-protein-coupled receptor, 2” 204287_at synaptogyrin 1 SYNGR1 1.63 3E−05 204161_s_at ectonucleotide pyrophosphatase/phosphodiesterase 4 ENPP4 1.62 2E−06 (putative function) 229000_at zinc finger protein 77 (pT1) ZNF77 1.62 4E−05 222423_at Nedd4 family interacting protein 1 NDFIP1 1.62 2E−14 204037_at “endothelial differentiation, lysophosphatidic acid EDG2 1.62 3E−09 G-protein-coupled receptor, 2” 219188_s_at LRP16 protein LRP16 1.62 5E−09 205404_at hydroxysteroid (11-beta) dehydrogenase 1 HSD11B1 1.62 1E−05 226959_at NA NA 1.62 3E−05 206176_at bone morphogenetic protein 6 BMP6 1.62 2E−06 1552789_at hypothetical protein FLJ32803 FLJ32803 1.62 1E−08 1558680_s_at “phosphodiesterase 1A, calmodulin-dependent” PDE1A 1.61 4E−05 205407_at reversion-inducing-cysteine-rich protein with kazal motifs RECK 1.61 7E−08 219039_at “sema domain, immunoglobulin domain (Ig), transmembrane SEMA4C 1.61 2E−07 domain (TM) and short cytoplasmic domain, (semaphorin) 4C” 224975_at nuclear factor I/A NFIA 1.61 1E−08 226197_at “Transcribed locus, strongly similar to XP_496055.1 NA 1.61 2E−07 similar to p40 [Homo sapiens]” 226360_at zinc and ring finger 3 ZNRF3 1.61 4E−07 208396_s_at “phosphodiesterase 1A, calmodulin-dependent” PDE1A 1.61 1E−06 213364_s_at Sorting nexin 1 SNX1 1.61 1E−08 225990_at brother of CDO BOC 1.60 1E−05 200635_sat “protein tyrosine phosphatase, receptor type, F” PTPRF 1.60 3E−05 203510_at met proto-oncogene (hepatocyte growth factor receptor) MET 1.60 2E−06 226322_at ARG99 protein ARG99 1.60 3E−07 205168_at “discoidin domain receptor family, member 2” DDR2 1.60 1E−06 232746_at Chemokine orphan receptor 1 CMKOR1 1.59 1E−07 238669_at prostaglandin-endoperoxide synthase 1 (prostaglandin PTGS1 1.59 1E−06 G/H synthase and cyclooxygenase) 229974_at Ellis van Creveld syndrome 2 (limbin) EVC2 1.59 5E−08 223611_s_at ligand of numb-protein X LNX 1.59 9E−06 226402_at “cytochrome P450, family 2, subfamily U, polypeptide 1” CYP2U1 1.59 1E−08 207031_at bagpipe homeobox homolog 1 (Drosophila) BAPX1 1.59 2E−05 235849_at hypothetical protein MGC45780 MGC45780 1.59 3E−08 218656_s_at lipoma HMGIC fusion partner LHFP 1.59 7E−07 205225_at estrogen receptor 1 ESR1 1.58 1E−08 1553347_s_at “potassium voltage-gated channel, shaker-related KCNA6 1.58 9E−05 subfamily, member 6” 1553995_a_at “5′-nucleotidase, ecto (CD73)” NT5E 1.58 2E−05 219354_at hypothetical protein FLJ11078 FLJ11078 1.58 5E−07 223603_at zinc finger protein 179 ZNF179 1.58 2E−05 211959_at insulin-like growth factor binding protein 5 IGFBP5 1.58 2E−05 224990_at hypothetical protein LOC201895 LOC201895 1.58 3E−11 222890_at HSPC065 protein HSPC065 1.58 2E−07 213131_at olfactomedin 1 OLFM1 1.58 7E−09 212977_at chemokine orphan receptor 1 CMKOR1 1.58 3E−07 203762_s_at dynein 2 light intermediate chain D2LIC 1.57 2E−10 231969_at NA NA 1.57 3E−05 201566_x_at “inhibitor of DNA binding 2, dominant negative ID2 1.57 7E−06 helix-loop-helix protein” 202242_at transmembrane 4 superfamily member 2 TM4SF2 1.57 6E−07 214620_x_at peptidylglycine alpha-amidating monooxygenase PAM 1.57 2E−09 225274_at prenylcysteine oxidase 1 PCYOX1 1.56 2E−13 228728_at hypothetical protein FLJ21986 FLJ21986 1.56 3E−07 218824_at hypothetical protein FLJ10781 FLJ10781 1.56 5E−05 233547_x_at “phosphodiesterase 1A, calmodulin-dependent” PDE1A 1.56 6E−05 205083_at aldehyde oxidase 1 AOX1 1.56 1E−05 227326_at Transmembrane anchor protein 1 TMAP1 1.56 2E−06 201120_s_at progesterone receptor membrane component 1 PGRMC1 1.56 7E−08 207076_s_at argininosuccinate synthetase ASS 1.56 1E−10 214651_s_at homeo box A9 HOXA9 1.56 2E−06 236644_at ring finger protein 180 RNF180 1.56 6E−05 209209_s_at “pleckstrin homology domain containing, family C PLEKHC1 1.56 1E−06 (with FERM domain) member 1” 230231_at “CDNA: FLJ23131 fis, clone LNG08502” NA 1.56 3E−05 210973_s_at “fibroblast growth factor receptor 1 (fms-related FGFR1 1.56 4E−09 tyrosine kinase 2, Pfeiffer syndrome)” 227126_at Transcribed locus NA 1.55 5E−08 218285_s_at dehydrogenase/reductase (SDR family) member 6 DHRS6 1.55 4E−10 219038_at “zinc finger, CW type with coiled-coil domain 2” ZCWCC2 1.55 3E−08 202017_at “epoxide hydrolase 1, microsomal (xenobiotic)” EPHX1 1.55 1E−06 213512_at chromosome 14 open reading frame 79 C14orf79 1.55 5E−05 218062_x_at CDC42 effector protein (Rho GTPase binding) 4 CDC42EP4 1.55 4E−06 210665_at tissue factor pathway inhibitor (lipoprotein-associated TFPI 1.55 3E−05 coagulation inhibitor) 201983_s_at “epidermal growth factor receptor (erythroblastic EGFR 1.55 5E−07 leukemia viral (v-erb-b) oncogene homolog, avian)” 211596_s_at leucine-rich repeats and immunoglobulin-like domains 1 | LRIG1 1.55 9E−07 leucine-rich repeats and immunoglobulin-like domains 1 201468_s_at “NAD(P)H dehydrogenase, quinone 1” NQO1 1.55 9E−05 209522_s_at carnitine acetyltransferase CRAT 1.55 2E−06 203343_at UDP-glucose dehydrogenase UGDH 1.55 2E−07 228184_at dispatched homolog 1 (Drosophila) DISP1 1.55 7E−07 213093_at “protein kinase C, alpha” PRKCA 1.55 2E−05 235956_at KIAA1377 protein KIAA1377 1.54 3E−07 202457_s_at “protein phosphatase 3 (formerly 2B), catalytic subunit, PPP3CA 1.54 6E−10 alpha isoform (calcineurin A alpha)” 226632_at cytoglobin CYGB 1.54 3E−05 226571_s_at “Protein tyrosine phosphatase, receptor type, S” PTPRS 1.54 1E−08 205651_x_at Rap guanine nucleotide exchange factor (GEF) 4 RAPGEF4 1.54 5E−07 212848_s_at chromosome 9 open reading frame 3 C9orf3 1.54 5E−07 203464_s_at epsin 2 EPN2 1.54 9E−12 221045_s_at period homolog 3 (Drosophila) PER3 1.54 9E−06 215039_at NA NA 1.54 2E−05 227719_at NA NA 1.54 2E−05 212325_at KIAA1102 protein KIAA1102 1.54 6E−10 206007_at proteoglycan 4 PRG4 1.53 7E−05 213227_at progesterone receptor membrane component 2 PGRMC2 1.53 3E−08 204570_at cytochrome c oxidase subunit VIIa polypeptide 1 (muscle) COX7A1 1.53 4E−07 204395_s_at G protein-coupled receptor kinase 5 GRK5 1.53 2E−09 230624_at “solute carrier family 25, member 27” SLC25A27 1.53 1E−05 215945_s_at tripartite motif-containing 2 TRIM2 1.53 8E−07 225946_at Chromosome 12 open reading frame 2 C12orf2 1.53 7E−06 227188_at chromosome 21 open reading frame 63 C21orf63 1.53 7E−06 226382_at hypothetical protein LOC283070 LOC283070 1.53 1E−05 1552476_s_at “phospholipase C, delta 3” PLCD3 1.53 5E−06 218418_s_at ankyrin repeat domain 25 ANKRD25 1.53 2E−08 213169_at Clone TUA8 Cri-du-chat region mRNA NA 1.53 9E−05 233136_at “poly(A) binding protein, cytoplasmic 5” PABPC5 1.53 5E−06 202328_s_at polycystic kidney disease 1 (autosomal dominant) PKD1 1.52 3E−07 226375_at Lemur tyrosine kinase 2 LMTK2 1.52 1E−05 230246_at placenta-specific 9 PLAC9 1.52 6E−05 227308_x_at latent transforming growth factor beta binding protein 3 LTBP3 1.52 3E−12 207895_at N-acetylated alpha-linked acidic dipeptidase-like 1 NAALADL1 1.52 1E−05 203803_at prenylcysteine oxidase 1 PCYOX1 1.52 6E−10 1553994_at “5′-nucleotidase, ecto (CD73)” NT5E 1.51 8E−05 212719_at “pleckstrin homology domain containing, family E PLEKHE1 1.51 5E−09 (with leucine rich repeats) member 1” 226931_at ARG99 protein ARG99 1.51 6E−06 202916_s_at “family with sequence similarity 20, member B” FAM20B 1.51 8E−12 226380_at “Protein tyrosine phosphatase, non-receptor type 21” PTPN21 1.51 6E−07 225868_at tripartite motif-containing 47 TRIM47 1.51 6E−05 202723_s_at forkhead box O1A (rhabdomyosarcoma) FOXO1A 1.51 2E−06 1553682_at F-box and leucine-rich repeat protein 14 FBXL14 1.50 6E−05 224973_at “family with sequence similarity 46, member A” FAM46A 1.50 4E−07 242033_at ring finger protein 180 RNF180 1.50 4E−05 212239_at “phosphoinositide-3-kinase, regulatory subunit 1 (p85 alpha)” PIK3R1 1.50 3E−07

TABLE 8 Gene expression associated with the F1 subtype of RA. NETAFFX: Fold ProbeID NETAFFX: GeneTitle GeneSymbol Change p-value 204848_x_at “hemoglobin, gamma A | hemoglobin, gamma A | HBG1 | 8.93 4E−06 hemoglobin, gamma G | hemoglobin, gamma G” HBG2 205959_at matrix metalloproteinase 13 (collagenase 3) | matrix metalloproteinase 13 (collagenase 3) MMP13 8.89 2E−09 213790_at A disintegrin and metalloproteinase domain 12 ADAM12 6.77 4E−12 (meltrin alpha) 228165_at hypothetical protein DKFZp547D2210 DKFZp547D2210 4.76 6E−07 205523_at hyaluronan and proteoglycan link protein 1 HAPLN1 4.74 6E−05 228703_at “procollagen-proline, 2-oxoglutarate 4-dioxygenase P4HA3 4.74 7E−20 (proline 4-hydroxylase), alpha polypeptide III” 202952_s_at a disintegrin and metalloproteinase domain 12 ADAM12 4.58 1E−11 (meltrin alpha) 204419_x_at “hemoglobin, gamma G | hemoglobin, gamma G” HBG2 4.21 6E−05 228640_at BH-protocadherin (brain-heart) PCDH7 4.20 5E−16 227372_s_at BAI1-associated protein 2-like 1 BAIAP2L1 4.10 1E−11 205902_at “potassium intermediate/small conductance KCNN3 4.07 2E−08 calcium-activated channel, subfamily N, member 3” 203798_s_at visinin-like 1 VSNL1 4.06 7E−16 219454_at “EGF-like-domain, multiple 6” EGFL6 3.86 5E−13 222862_s_at adenylate kinase 5 AK5 3.75 1E−07 203936_s_at “matrix metalloproteinase 9 (gelatinase B, 92 kDa MMP9 3.74 2E−06 gelatinase, 92 kDa type IV collagenase)” 223721_s_at “DnaJ (Hsp40) homolog, subfamily C, member 12” DNAJC12 3.73 2E−11 230895_at Hyaluronan and proteoglycan link protein 1 HAPLN1 3.54 4E−05 213425_at “wingless-type MMTV integration site family, member 5A | WNT5A 3.47 1E−13 wingless-type MMTV integration site family, member 5A” 1554863_s_at docking protein 5 DOK5 3.45 1E−05 223121_s_at secreted frizzled-related protein 2 SFRP2 3.42 5E−07 207118_s_at matrix metalloproteinase 23B | MMP23B | 3.39 6E−05 matrix metalloproteinase 23A MMP23A 204379_s_at “fibroblast growth factor receptor 3 (achondroplasia, FGFR3 3.34 2E−08 thanatophoric dwarfism)” 232122_s_at NA NA 3.07 6E−11 227860_at carboxypeptidase X (M14 family) CPXM 3.06 2E−09 223722_at “DnaJ (Hsp40) homolog, subfamily C, member 12” DNAJC12 3.04 3E−07 230204_at Hyaluronan and proteoglycan link protein 1 HAPLN1 3.03 4E−05 230464_at “endothelial differentiation, sphingolipid EDG8 2.92 5E−12 G-protein-coupled receptor, 8” 210134_x_at short stature homeobox 2 SHOX2 2.91 1E−07 202935_s_at “SRY (sex determining region Y)-box 9 (campomelic SOX9 2.91 1E−07 dysplasia, autosomal sex-reversal)” 213059_at cAMP responsive element binding protein 3-like 1 CREB3L1 2.91 3E−19 205990_s_at “wingless-type MMTV integration site family, WNT5A 2.90 2E−13 member 5A” 225681_at collagen triple helix repeat containing 1 CTHRC1 2.89 3E−12 1554697_at “a disintegrin-like and metalloprotease (reprolysin type) ADAMTS9 2.89 8E−08 with thrombospondin type 1 motif, 9” 221697_at MAP1 light chain 3-like protein 2 | LOC440738 2.88 4E−05 MAP1 light chain 3-like protein 2 203184_at fibrillin 2 (congenital contractural arachnodactyly) FBN2 2.84 4E−07 218976_at “DnaJ (Hsp40) homolog, subfamily C, member 12” DNAJC12 2.83 1E−11 212489_at “Collagen, type V, alpha 1” COL5A1 2.83 5E−13 209800_at keratin 16 (focal non-epidermolytic palmoplantar KRT16 2.83 8E−05 keratoderma) 205524_s_at hyaluronan and proteoglycan link protein 1 HAPLN1 2.78 9E−05 226814_at “a disintegrin-like and metalloprotease (reprolysin type) ADAMTS9 2.76 1E−08 with thrombospondin type 1 motif, 9” 206933_s_at hexose-6-phosphate dehydrogenase H6PD 2.76 1E−10 (glucose 1-dehydrogenase) 205131_x_at “C-type lectin domain family 11, member A” CLEC11A 2.75 1E−12 205375_at MyoD family inhibitor MDFI 2.74 6E−12 236044_at phosphatidic acid phosphatase type 2 domain containing 1 PPAPDC1 2.72 9E−12 1554293_at tau tubulin kinase 2 TTBK2 2.72 7E−08 215271_at tenascin N TNN 2.72 3E−05 206842_at “potassium voltage-gated channel, Shal-related KCND1 2.71 1E−06 subfamily, member 1” 227971_at Nik related kinase NRK 2.70 2E−11 206376_at “solute carrier family 6 (neurotransmitter transporter), SLC6A15 2.68 3E−06 member 15” 205381_at leucine rich repeat containing 17 LRRC17 2.67 8E−12 211396_at “Fc fragment of IgG, low affinity IIc, receptor for (CD32)” FCGR2C 2.67 5E−08 224941_at “pregnancy-associated plasma protein A, pappalysin 1” PAPPA 2.64 4E−05 209035_at midkine (neurite growth-promoting factor 2) MDK 2.64 6E−09 210151_s_at dual-specificity tyrosine-(Y)-phosphorylation DYRK3 2.62 1E−07 regulated kinase 3 1555256_at Ellis van Creveld syndrome 2 (limbin) EVC2 2.62 4E−09 204358_s_at fibronectin leucine rich transmembrane protein 2 FLRT2 2.59 6E−14 219894_at MAGE-like 2 MAGEL2 2.59 5E−10 236258_at chromosome 20 open reading frame 151 C20orf151 2.57 5E−06 204359_at fibronectin leucine rich transmembrane protein 2 FLRT2 2.57 5E−13 235510_at Usher syndrome 1C binding protein 1 USHBP1 2.56 9E−07 233030_at adiponutrin ADPN 2.56 5E−06 203325_s_at “collagen, type V, alpha 1” COL5A1 2.56 2E−12 204281_at TEA domain family member 4 TEAD4 2.54 2E−09 224207_x_at matrix metalloproteinase 28 MMP28 2.53 3E−09 215446_s_at lysyl oxidase LOX 2.52 1E−09 212473_s_at flavoprotein oxidoreductase MICAL2 MICAL2 2.52 1E−09 205866_at ficolin (collagen/fibrinogen domain containing) 3 FCN3 2.52 6E−05 (Hakata antigen) 211175_at G protein-coupled receptor 45 GPR45 2.51 9E−10 201107_s_at thrombospondin 1 THBS1 2.50 6E−05 1553027_a_at kelch-like 4 (Drosophila) KLHL4 2.49 3E−06 205535_s_at BH-protocadherin (brain-heart) PCDH7 2.49 4E−08 211899_s_at TNF receptor-associated factor 4 TRAF4 2.48 2E−06 236179_at “Cadherin 11, type 2, OB-cadherin (osteoblast)” CDH11 2.48 1E−13 200884_at “creatine kinase, brain” CKB 2.48 3E−06 236245_at outer dense fiber of sperm tails 3-like 1 ODF3L1 2.46 3E−05 223122_s_at secreted frizzled-related protein 2 SFRP2 2.45 4E−07 211062_s_at carboxypeptidase Z | carboxypeptidase Z CPZ 2.45 2E−06 204123_at “ligase III, DNA, ATP-dependent” LIG3 2.44 2E−10 219263_at ring finger protein 128 RNF128 2.43 6E−07 219555_s_at uncharacterized bone marrow protein BM039 BM039 2.42 1E−05 213155_at KIAA0523 protein KIAA0523 2.42 1E−05 217430_x_at NA NA 2.41 2E−10 228367_at alpha-kinase 2 ALPK2 2.41 2E−10 230240_at Transcribed locus NA 2.40 2E−08 205656_at protocadherin 17 PCDH17 2.40 7E−05 220289_s_at absent in melanoma 1-like AIM1L 2.40 2E−05 213085_s_at KIBRA protein KIBRA 2.40 9E−08 205031_at ephrin-B3 EFNB3 2.39 3E−05 206234_s_at matrix metalloproteinase 17 (membrane-inserted) MMP17 2.39 2E−06 206655_s_at “glycoprotein Ib (platelet), beta polypeptide” GP1BB 2.38 2E−06 218051_s_at hypothetical protein FLJ12442 FLJ12442 2.37 3E−11 203823_at regulator of G-protein signalling 3 RGS3 2.37 1E−09 218653_at solute carrier family 25 (mitochondrial carrier; SLC25A15 2.36 2E−06 ornithine transporter) member 15 207714_s_at “serine (or cysteine) proteinase inhibitor, clade H (heat shock SERPINH1 2.36 3E−12 protein 47), member 1, (collagen binding protein 1)” 202148_s_at pyrroline-5-carboxylate reductase 1 PYCR1 2.35 3E−17 204600_at EPH receptor B3 EPHB3 2.35 1E−11 219602_s_at “family with sequence similarity 38, member B” FAM38B 2.34 8E−08 213170_at glutathione peroxidase 7 GPX7 2.34 4E−14 210477_x_at mitogen-activated protein kinase 8 MAPK8 2.34 1E−08 239542_at NA NA 2.32 7E−06 227289_at hypothetical protein LOC144997 LOC144997 2.31 6E−05 202311_s_at “collagen, type I, alpha 1” COL1A1 2.31 1E−08 210323_at tektin 2 (testicular) TEKT2 2.31 4E−05 214608_s_at eyes absent homolog 1 (Drosophila) EYA1 2.31 2E−06 204904_at “gap junction protein, alpha 4, 37 kDa (connexin 37)” GJA4 2.30 2E−08 204826_at cyclin F CCNF 2.30 8E−07 211233_x_at estrogen receptor 1 ESR1 2.30 1E−08 206201_s_at mesenchyme homeo box 2 (growth arrest-specific homeo box) MEOX2 2.29 3E−07 221730_at “collagen, type V, alpha 2” COL5A2 2.29 2E−12 235545_at DEP domain containing 1 DEPDC1 2.29 2E−05 207379_at EGF-like repeats and discoidin I-like domains 3 EDIL3 2.29 4E−08 220979_s_at “ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N- ST6GALNAC5 2.28 6E−08 acetylgalactosaminide alpha-2,6-sialyltransferase 5 | ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N- acetylgalactosaminide alpha-2,6-sialyltransferase 5” 244533_at NA NA 2.28 3E−08 220093_at anthrax toxin receptor 1 ANTXR1 2.28 8E−05 235944_at hemicentin FIBL-6 2.27 2E−10 233026_s_at PDZ domain containing 3 PDZK3 2.27 3E−07 200644_at MARCKS-like 1 MARCKSL1 2.26 6E−08 203434_s_at “membrane metallo-endopeptidase (neutral endopeptidase, MME 2.26 6E−07 enkephalinase, CALLA, CD10)” 228128_x_at “pregnancy-associated plasma protein A, pappalysin 1” PAPPA 2.25 2E−06 205262_at “potassium voltage-gated channel, subfamily H (eag-related), KCNH2 2.25 1E−10 member 2” 213869_x_at Thy-1 cell surface antigen | Thy-1 co-transcribed THY1 | LOC94105 2.25 8E−10 206547_s_at “protein phosphatase, EF hand calcium-binding domain 1” PPEF1 2.24 1E−07 204078_at synaptonemal complex protein SC65 SC65 2.24 2E−14 238732_at “collagen, type XXIV, alpha 1” COL24A1 2.23 7E−11 236689_at ring finger protein 151 RNF151 2.23 2E−07 206528_at “transient receptor potential cation channel, subfamily C, TRPC6 2.23 3E−13 member 6” 243502_at “Gap junction protein, alpha 7, 45 kDa (connexin 45)” GJA7 2.22 4E−10 223574_x_at “protein phosphatase 2 (formerly 2A), regulatory subunit B (PR 52), gamma isoform” PPP2R2C 2.22 5E−07 223587_s_at amnionless homolog (mouse) AMN 2.22 1E−08 218638_s_at “spondin 2, extracellular matrix protein” SPON2 2.22 5E−07 219401_at xylosyltransferase II XYLT2 2.21 1E−07 1557176_a_at chromosome 14 open reading frame 37 C14orf37 2.20 5E−11 204298_s_at lysyl oxidase LOX 2.19 7E−08 206754_s_at “cytochrome P450, family 2, subfamily B, polypeptide 6” CYP2B6 2.19 8E−06 228814_at retinoblastoma binding protein 6 RBBP6 2.19 2E−08 209031_at “Immunoglobulin superfamily, member 4” IGSF4 2.18 3E−06 1438_at EPH receptor B3 EPHB3 2.18 2E−12 227557_at “scavenger receptor class F, member 2” SCARF2 2.17 3E−08 211071_s_at ALL1-fused gene from chromosome 1q | AF1Q 2.17 1E−09 ALL1-fused gene from chromosome 1q 210095_s_at insulin-like growth factor binding protein 3 IGFBP3 2.17 3E−07 206758_at endothelin 2 EDN2 2.17 6E−05 1558643_s_at EGF-like repeats and discoidin I-like domains 3 EDIL3 2.16 1E−11 234605_at CDC14 cell division cycle 14 homolog B (S. cerevisiae) CDC14B 2.16 6E−05 226891_at chromosome 3 open reading frame 21 C3orf21 2.15 8E−05 235343_at Hypothetical protein FLJ12505 FLJ12505 2.15 7E−06 202936_s_at “SRY (sex determining region Y)-box 9 (campomelic SOX9 2.15 2E−06 dysplasia, autosomal sex-reversal)” 228080_at layilin LOC143903 2.14 2E−13 213640_s_at lysyl oxidase LOX 2.14 2E−05 204941_s_at “aldehyde dehydrogenase 3 family, member B2” ALDH3B2 2.14 8E−05 218898_at membrane protein expressed in epithelial-like CT120 2.13 2E−17 lung adenocarcinoma 222461_s_at hect domain and RLD 2 HERC2 2.13 1E−06 229942_at Transcribed locus NA 2.13 3E−11 201981_at “pregnancy-associated plasma protein A, pappalysin 1” PAPPA 2.12 2E−08 204040_at ring finger protein 144 RNF144 2.11 4E−12 1561853_a_at interleukin 23 receptor IL23R 2.11 7E−05 205612_at multimerin 1 MMRN1 2.11 2E−05 211709_s_at “C-type lectin domain family 11, member A | CLEC11A 2.09 4E−11 C-type lectin domain family 11, member A” 209082_s_at “collagen, type XVIII, alpha 1” COL18A1 2.09 4E−13 210783_x_at “C-type lectin domain family 11, member A” CLEC11A 2.08 4E−06 211685_s_at neurocalcin delta | neurocalcin delta NCALD 2.08 2E−11 202007_at nidogen (enactin) NID 2.08 3E−11 226769_at similar to RIKEN cDNA 1110018M03 LOC387758 2.07 1E−05 219310_at chromosome 20 open reading frame 39 C20orf39 2.07 4E−06 242100_at chondroitin sulfate synthase 3 CSS3 2.07 4E−10 213695_at paraoxonase 3 PON3 2.06 5E−05 223170_at DKFZP564K1964 protein DKFZP564K1964 2.06 1E−10 213707_s_at distal-less homeo box 5 DLX5 2.05 2E−05 204464_s_at endothelin receptor type A EDNRA 2.05 5E−13 1554989_at KIAA0317 KIAA0317 2.04 5E−05 203636_at midline 1 (Opitz/BBB syndrome) MID1 2.04 5E−12 204463_s_at endothelin receptor type A EDNRA 2.03 1E−07 243409_at Forkhead box L1 FOXL1 2.03 4E−05 1553889_at G protein-coupled receptor MRGX2 MRGX2 2.03 9E−05 227314_at “Integrin, alpha 2 (CD49B, alpha 2 subunit of ITGA2 2.03 3E−11 VLA-2 receptor)” 221729_at “collagen, type V, alpha 2” COL5A2 2.02 2E−11 214234_s_at “cytochrome P450, family 3, subfamily A, CYP3A5 2.02 1E−04 polypeptide 5” 204468_s_at tyrosine kinase with immunoglobulin-like and TIE1 2.02 3E−07 EGF-like domains 1 223235_s_at SPARC related modular calcium binding 2 SMOC2 2.02 1E−10 202409_at putative insulin-like growth factor II associated protein LOC492304 2.01 2E−05 238805_at similar to RIKEN cDNA 2310030G06 gene MGC14839 2.01 1E−06 208224_at homeo box B1 HOXB1 2.01 4E−09 208850_s_at Thy-1 cell surface antigen | Thy-1 co-transcribed THY1 | LOC94105 2.01 2E−09 202894_at EPH receptor B4 EPHB4 2.01 2E−10 239286_at “Cadherin 11, type 2, OB-cadherin (osteoblast)” CDH11 2.00 2E−09 202068_s_at low density lipoprotein receptor LDLR 2.00 3E−05 (familial hypercholesterolemia) 1558342_x_at DIX domain containing 1 DIXDC1 2.00 1E−08 206106_at mitogen-activated protein kinase 12 MAPK12 2.00 1E−09 218730_s_at “osteoglycin (osteoinductive factor, mimecan)” OGN 2.00 4E−06 207039_at “cyclin-dependent kinase inhibitor 2A CDKN2A 2.00 8E−05 (melanoma, p16, inhibits CDK4)” 205226_at platelet-derived growth factor receptor-like PDGFRL 1.99 6E−07 202067_s_at low density lipoprotein receptor LDLR 1.99 3E−05 (familial hypercholesterolemia) 218717_s_at leprecan-like 1 LEPREL1 1.99 4E−09 209081_s_at “collagen, type XVIII, alpha 1” COL18A1 1.99 4E−14 215125_s_at “UDP glycosyltransferase 1 family, polypeptide A10 | UGT1A10 | 1.99 2E−05 UDP glycosyltransferase 1 family, polypeptide A8 | UGT1A8 | UDP glycosyltransferase 1 family, polypeptide A7 | UGT1A7 | UDP glycosyltransferase 1 family, polypeptide A6 | UGT1A6 | UDP glycosyltransferase 1 family, polypeptide . . .” UGT1A5 | UGT1A9 | UGT1A4 | UGT1A1 | UGT1A3 219686_at serine/threonine kinase 32B STK32B 1.98 2E−06 211234_x_at estrogen receptor 1 ESR1 1.98 3E−07 1564746_at hypothetical protein BC009732 LOC133308 1.97 1E−07 238169_at Transcribed locus NA 1.97 9E−09 203780_at epithelial V-like antigen 1 EVA1 1.96 4E−06 218629_at smoothened homolog (Drosophila) SMO 1.96 5E−05 219102_at “reticulocalbin 3, EF-hand calcium binding domain” RCN3 1.96 7E−11 209841_s_at leucine rich repeat neuronal 3 LRRN3 1.96 1E−04 211343_s_at “collagen, type XIII, alpha 1” COL13A1 1.96 2E−07 214347_s_at dopa decarboxylase (aromatic L-amino acid decarboxylase) DDC 1.96 1E−06 203637_s_at midline 1 (Opitz/BBB syndrome) MID1 1.94 2E−12 222856_at “apelin, AGTRL1 ligand” APLN 1.94 8E−06 207011_s_at PTK7 protein tyrosine kinase 7 PTK7 1.94 1E−13 219419_at chromosome 18 open reading frame 22 C18orf22 1.93 2E−05 222548_s_at mitogen-activated protein kinase kinase kinase kinase 4 MAP4K4 1.92 3E−15 219556_at hypothetical protein FLJ13909 FLJ13909 1.92 1E−07 228776_at “Gap junction protein, alpha 7, 45 kDa (connexin 45)” GJA7 1.91 3E−11 209427_at smoothelin SMTN 1.91 2E−10 220807_at “hemoglobin, theta 1 | hemoglobin, theta 1” HBQ1 1.90 7E−05 1554398_at lysozyme-like LYG2 1.90 9E−05 208370_s_at Down syndrome critical region gene 1 DSCR1 1.90 3E−06 213435_at SATB family member 2 SATB2 1.90 1E−06 226244_at “C-type lectin domain family 14, member A” CLEC14A 1.90 1E−06 209812_x_at “caspase 2, apoptosis-related cysteine protease CASP2 1.90 2E−06 (neural precursor cell expressed, developmentally down-regulated 2)” 227204_at par-6 partitioning defective 6 homolog gamma (C. elegans) PARD6G 1.90 2E−11 223594_at hypothetical protein DKFZp434K2435 DKFZp434K2435 1.90 6E−09 227468_at carnitine palmitoyltransferase 1C CPT1C 1.89 4E−08 1557000_at hypothetical protein LOC339768 LOC339768 1.89 5E−11 225790_at methionine sulfoxide reductase B3 MSRB3 1.89 6E−09 232566_at nucleolar protein family 6 (RNA-associated) NOL6 1.88 3E−09 213125_at olfactomedin-like 2B OLFML2B 1.88 3E−09 200907_s_at palladin KIAA0992 1.88 6E−09 206775_at cubilin (intrinsic factor-cobalamin receptor) CUBN 1.88 3E−05 1552672_a_at “immunoglobulin superfamily, member 3” IGSF3 1.88 5E−07 1559394_a_at Receptor tyrosine kinase-like orphan receptor 1 ROR1 1.87 2E−06 230722_at NA NA 1.87 1E−13 219390_at “FK506 binding protein 14, 22 kDa” FKBP14 1.87 1E−11 212558_at “sprouty homolog 1, antagonist of FGF signaling (Drosophila)” SPRY1 1.87 3E−05 211140_s_at “caspase 2, apoptosis-related cysteine protease CASP2 1.87 1E−09 (neural precursor cell expressed, developmentally down-regulated 2)” 61734_at “reticulocalbin 3, EF-hand calcium binding domain” RCN3 1.87 4E−11 206571_s_at mitogen-activated protein kinase kinase kinase kinase 4 MAP4K4 1.86 3E−10 1562528_at RAR-related orphan receptor A RORA 1.86 8E−06 227850_x_at CDC42 effector protein (Rho GTPase binding) 5 CDC42EP5 1.86 8E−10 202112_at von Willebrand factor VWF 1.86 1E−08 209890_at transmembrane 4 superfamily member 9 | TM4SF9 1.86 6E−10 transmembrane 4 superfamily member 9 207173_x_at “cadherin 11, type 2, OB-cadherin (osteoblast)” CDH11 1.85 1E−11 213139_at snail homolog 2 (Drosophila) SNAI2 1.85 7E−08 226960_at DMC UNQ473 1.84 3E−05 220226_at “transient receptor potential cation channel, TRPM8 1.84 3E−05 subfamily M, member 8” 204141_at “tubulin, beta 2” TUBB2 1.84 1E−07 209030_s_at “immunoglobulin superfamily, member 4” IGSF4 1.84 1E−05 40016_g_at microtubule associated serine/threonine kinase MAST4 1.83 3E−06 family member 4 40687_at “gap junction protein, alpha 4, 37 kDa (connexin 37)” GJA4 1.83 4E−07 234610_at heat shock 70 kD protein 12B HSPA12B 1.83 4E−08 204017_at KDEL (Lys-Asp-Glu-Leu) endoplasmic reticulum protein KDELR3 1.83 1E−10 retention receptor 3 204901_at beta-transducin repeat containing BTRC 1.83 1E−05 206116_s_at tropomyosin 1 (alpha) TPM1 1.82 2E−09 208851_s_at Thy-1 cell surface antigen | Thy-1 co-transcribed THY1 | LOC94105 1.82 5E−10 205289_at bone morphogenetic protein 2 BMP2 1.82 1E−05 201445_at “calponin 3, acidic” CNN3 1.82 1E−05 206310_at “serine protease inhibitor, Kazal type 2 SPINK2 1.82 3E−05 (acrosin-trypsin inhibitor)” 242671_at Midline 1 (Opitz/BBB syndrome) MID1 1.81 2E−07 218181_s_at mitogen-activated protein kinase kinase kinase kinase 4 MAP4K4 1.81 3E−12 205578_at receptor tyrosine kinase-like orphan receptor 2 ROR2 1.81 3E−07 229369_at V-set and immunoglobulin domain containing 2 VSIG2 1.81 2E−06 214043_at “Protein tyrosine phosphatase, receptor type, D” PTPRD 1.81 2E−06 207172_s_at “cadherin 11, type 2, OB-cadherin (osteoblast)” CDH11 1.80 1E−10 213345_at “nuclear factor of activated T-cells, cytoplasmic, NFATC4 1.80 8E−07 calcineurin-dependent 4” 201792_at AE binding protein 1 AEBP1 1.80 7E−07 224817_at SH3 multiple domains 1 SH3MD1 1.80 3E−10 228748_at “CD59 antigen p18-20 (antigen identified by monoclonal CD59 1.79 1E−08 antibodies 16.3A5, EJ16, EJ30, EL32 and G344)” 213362_at “Protein tyrosine phosphatase, receptor type, D” PTPRD 1.79 4E−06 200906_s_at palladin KIAA0992 1.79 5E−07 201416_at SRY (sex determining region Y)-box 4 SOX4 1.79 8E−05 234994_at KIAA1913 KIAA1913 1.79 6E−10 209655_s_at transmembrane 4 superfamily member 10 TM4SF10 1.79 5E−05 205122_at transmembrane protein with EGF-like and two TMEFF1 1.78 6E−09 follistatin-like domains 1 212915_at PDZ domain containing RING finger 3 PDZRN3 1.78 7E−07 217287_s_at “transient receptor potential cation channel, subfamily C, TRPC6 1.78 3E−08 member 6” 242979_at Transcribed locus NA 1.77 1E−08 205453_at homeo box B2 HOXB2 1.77 8E−07 233365_at “Cleavage stimulation factor, 3′ pre-RNA, subunit 2, CSTF2T 1.77 5E−05 64 kDa, tau variant” 213496_at plasticity related gene 1 LPPR4 1.77 5E−05 210330_at “sarcoglycan, delta (35 kDa dystrophin-associated glycoprotein)” SGCD 1.76 3E−10 204115_at “guanine nucleotide binding protein (G protein), gamma 11” GNG11 1.76 2E−09 214319_at Hypothetical protein CG003 13CDNA73 1.76 2E−06 220014_at mesenchymal stem cell protein DSC54 LOC51334 1.76 4E−05 225782_at methionine sulfoxide reductase B3 MSRB3 1.75 2E−07 204686_at insulin receptor substrate 1 IRS1 1.75 2E−08 218834_s_at “heat shock 70 kDa protein 5 (glucose-regulated protein, 78 kDa) binding protein 1” HSPA5BP1 1.75 2E−06 202008_s_at nidogen (enactin) NID 1.75 6E−09 209656_s_at transmembrane 4 superfamily member 10 TM4SF10 1.75 8E−06 202450_s_at cathepsin K (pycnodysostosis) CTSK 1.75 4E−14 213252_at SH3 multiple domains 1 SH3MD1 1.75 3E−09 209561_at thrombospondin 3 THBS3 1.74 5E−06 204688_at “sarcoglycan, epsilon” SGCE 1.74 3E−06 205303_at “potassium inwardly-rectifying channel, subfamily J, member 8” KCNJ8 1.74 2E−08 228054_at transmembrane protein 44 TMEM44 1.74 4E−05 228067_at similar to 2010300C02Rik protein MGC42367 1.73 3E−06 202503_s_at KIAA0101 KIAA0101 1.73 5E−05 219637_at hypothetical protein FLJ12584 FLJ12584 1.73 1E−06 238478_at Basonuclin 2 BNC2 1.73 1E−08 220272_at basonuclin 2 BNC2 1.73 1E−09 202871_at TNF receptor-associated factor 4 TRAF4 1.73 7E−08 201505_at “laminin, beta 1” LAMB1 1.73 1E−08 229172_at heat shock 70 kD protein 12B HSPA12B 1.72 3E−07 225735_at KIAA1223 protein KIAA1223 1.72 3E−09 217791_s_at “aldehyde dehydrogenase 18 family, member A1” ALDH18A1 1.72 2E−11 41037_at TEA domain family member 4 TEAD4 1.72 3E−07 217963_s_at nerve growth factor receptor (TNFRSF16) associated protein 1 NGFRAP1 1.72 3E−08 225611_at microtubule associated serine/threonine kinase family member 4 MAST4 1.71 3E−06 212533_at WEE1 homolog (S. pombe) WEE1 1.71 3E−07 221870_at EH-domain containing 2 EHD2 1.71 5E−07 218839_at hairy/enhancer-of-split related with YRPW motif 1 HEY1 1.71 2E−08 219134_at “EGF, latrophilin and seven transmembrane domain containing 1” ELTD1 1.71 4E−08 238583_at methionine sulfoxide reductase B3 MSRB3 1.71 2E−06 202712_s_at “creatine kinase, mitochondrial 1 (ubiquitous)” CKMT1 1.71 3E−06 212651_at Rho-related BTB domain containing 1 RHOBTB1 1.71 4E−11 209633_at “protein phosphatase 2 (formerly 2A), regulatory subunit B″, alpha” PPP2R3A 1.70 2E−05 1553620_at tripartite motif-containing 42 TRIM42 1.70 5E−06 1568618_a_at UDP-N-acetyl-alpha-D-galactosamine: polypeptide N- GALNT1 1.70 2E−09 acetylgalactosaminyltransferase 1 (GalNAc-T1) 226282_at Full length insert cDNA clone ZE03F06 NA 1.70 8E−05 206607_at Cas-Br-M (murine) ecotropic retroviral transforming sequence CBL 1.70 2E−06 212364_at myosin IB MYO1B 1.69 1E−09 242668_x_at sperm associated antigen 4-like SPAG4L 1.69 3E−05 209694_at 6-pyruvoyltetrahydropterin synthase PTS 1.69 1E−11 202949_s_at four and a half LIM domains 2 FHL2 1.69 2E−07 212950_at G protein-coupled receptor 116 GPR116 1.69 2E−09 212365_at myosin IB MYO1B 1.69 7E−11 1562529_s_at RAR-related orphan receptor A RORA 1.69 9E−05 208664_s_at tetratricopeptide repeat domain 3 TTC3 1.68 3E−07 222722_at “osteoglycin (osteoinductive factor, mimecan)” OGN 1.68 1E−05 214844_s_at docking protein 5 DOK5 1.68 6E−05 224942_at “pregnancy-associated plasma protein A, pappalysin 1” PAPPA 1.68 8E−05 228563_at NA NA 1.68 6E−09 230972_at ankyrin repeat domain 9 ANKRD9 1.68 2E−06 226905_at hypothetical protein MGC45871 MGC45871 1.68 9E−06 204136_at “collagen, type VII, alpha 1 (epidermolysis bullosa, dystrophic, COL7A1 1.67 1E−06 dominant and recessive)” 221401_at “calcium channel, voltage-dependent, gamma subunit 5” CACNG5 1.67 1E−05 222101_s_at dachsous 1 (Drosophila) DCHS1 1.67 5E−08 231175_at chromosome 6 open reading frame 65 C6orf65 1.67 4E−07 222675_s_at BAI1-associated protein 2-like 1 BAIAP2L1 1.67 2E−07 207390_s_at smoothelin SMTN 1.67 5E−08 204983_s_at glypican 4 GPC4 1.67 4E−06 203150_at Rab9 effector p40 RAB9P40 1.67 5E−09 33579_i_at galanin receptor 3 GALR3 1.67 5E−06 200897_s_at palladin KIAA0992 1.66 4E−08 219522_at four jointed box 1 (Drosophila) FJX1 1.66 2E−06 210987_x_at Tropomyosin 1 (alpha) TPM1 1.66 2E−07 205304_s_at “potassium inwardly-rectifying channel, subfamily J, KCNJ8 1.66 2E−07 member 8” 208663_s_at tetratricopeptide repeat domain 3 TTC3 1.66 4E−10 225322_s_at hypothetical protein FLJ22175 FLJ22175 1.66 1E−04 206702_at “TEK tyrosine kinase, endothelial (venous malformations, TEK 1.66 3E−06 multiple cutaneous and mucosal)” 212642_s_at human immunodeficiency virus type I enhancer HIVEP2 1.66 2E−05 binding protein 2 208661_s_at tetratricopeptide repeat domain 3 TTC3 1.66 7E−13 228396_at NA NA 1.66 2E−07 226893_at “V-abl Abelson murine leukemia viral oncogene homolog 2 ABL2 1.65 3E−05 (arg, Abelson-related gene)” 201681_s_at “discs, large homolog 5 (Drosophila)” DLG5 1.65 5E−06 221024_s_at “solute carrier family 2 (facilitated glucose transporter), SLC2A10 1.65 2E−08 member 10 | solute carrier family 2 (facilitated glucose transporter), member 10” 201261_x_at biglycan BGN 1.65 2E−05 205104_at syntaphilin SNPH 1.65 3E−06 224998_at chemokine-like factor super family 4 CKLFSF4 1.65 3E−09 225736_at F-box protein 22 FBXO22 1.65 4E−05 226876_at hypothetical protein MGC45871 MGC45871 1.65 7E−06 226950_at Activin A receptor type II-like 1 ACVRL1 1.65 6E−05 222416_at “aldehyde dehydrogenase 18 family, member A1” ALDH18A1 1.65 7E−12 237411_at “a disintegrin-like and metalloprotease (reprolysin type) with ADAMTS6 1.64 6E−05 thrombospondin type 1 motif, 6” 225613_at microtubule associated serine/threonine kinase family member 4 MAST4 1.64 3E−07 207119_at “protein kinase, cGMP-dependent, type I” PRKG1 1.64 1E−06 208073_x_at tetratricopeptide repeat domain 3 TTC3 1.64 8E−14 224746_at KIAA1522 protein KIAA1522 1.64 4E−05 210135_s_at short stature homeobox 2 SHOX2 1.64 1E−05 204400_at embryonal Fyn-associated substrate EFS 1.64 2E−05 204966_at brain-specific angiogenesis inhibitor 2 BAI2 1.64 4E−06 219051_x_at “meteorin, glial cell differentiation regulator” METRN 1.64 2E−08 237929_at LOC146853 LOC146853 1.64 8E−05 201418_s_at SRY (sex determining region Y)-box 4 SOX4 1.64 1E−06 207264_at KDEL (Lys-Asp-Glu-Leu) endoplasmic reticulum protein KDELR3 1.64 5E−06 retention receptor 3 215074_at myosin IB MYO1B 1.63 2E−05 210645_s_at tetratricopeptide repeat domain 3 TTC3 1.63 2E−11 201724_s_at UDP-N-acetyl-alpha-D-galactosamine: polypeptide N- GALNT1 1.63 3E−09 acetylgalactosaminyltransferase 1 (GalNAc-T1) 209596_at adlican DKFZp564I1922 1.63 7E−06 210986_s_at tropomyosin 1 (alpha) TPM1 1.63 4E−07 205164_at glycine C-acetyltransferase (2-amino-3-ketobutyrate GCAT 1.63 4E−05 coenzyme A ligase) 200757_s_at calumenin CALU 1.62 5E−08 233025_at PDZ domain containing 3 PDZK3 1.62 2E−05 232269_x_at “meteorin, glial cell differentiation regulator” METRN 1.62 6E−09 227399_at Colon carcinoma related protein FLJ38507 1.62 1E−05 227481_at membrane associated guanylate kinase interacting MAGI1 1.62 4E−05 protein-like 1 44783_s_at hairy/enhancer-of-split related with YRPW motif 1 HEY1 1.62 5E−09 222937_s_at matrix metalloproteinase 28 MMP28 1.62 1E−06 235204_at “COX15 homolog, cytochrome c oxidase assembly protein (yeast)” COX15 1.61 3E−07 208682_s_at “melanoma antigen family D, 2” MAGED2 1.61 1E−08 232080_at “HECT, C2 and WW domain containing E3 ubiquitin HECW2 1.61 4E−05 protein ligase 2” 211651_s_at “laminin, beta 1 | laminin, beta 1” LAMB1 1.61 1E−07 213221_s_at SNF1-like kinase 2 SNF1LK2 1.61 7E−05 217312_s_at “collagen, type VII, alpha 1 (epidermolysis bullosa, COL7A1 1.61 9E−05 dystrophic, dominant and recessive)” 210869_s_at melanoma cell adhesion molecule MCAM 1.61 8E−05 225009_at chemokine-like factor super family 4 CKLFSF4 1.61 1E−08 222803_at phosphoribosyl transferase domain containing 1 PRTFDC1 1.61 9E−08 207265_s_at KDEL (Lys-Asp-Glu-Leu) endoplasmic reticulum protein KDELR3 1.60 5E−10 retention receptor 3 213905_x_at biglycan | serologically defined colon cancer antigen 33 BGN | SDCCAG33 1.60 5E−06 213012_at “neural precursor cell expressed, developmentally NEDD4 1.60 1E−09 down-regulated 4” 210958_s_at microtubule associated serine/threonine kinase family MAST4 1.60 8E−06 member 4 1568779_a_at “extracellular matrix protein 2, female organ and ECM2 1.60 6E−08 adipocyte specific” 204854_at leprecan-like 2 LEPREL2 1.60 4E−06 45297_at EH-domain containing 2 EHD2 1.60 4E−07 212641_at human immunodeficiency virus type I enhancer HIVEP2 1.60 3E−05 binding protein 2 221880_s_at hypothetical gene supported by AK075564; BC060873 LOC400451 1.59 1E−05 221599_at PTD015 protein PTD015 1.59 8E−05 215017_s_at formin binding protein 1-like FNBP1L 1.59 1E−06 216235_s_at endothelin receptor type A EDNRA 1.59 1E−06 210993_s_at “SMAD, mothers against DPP homolog 1 (Drosophila)” SMAD1 1.59 3E−05 207836_s_at RNA binding protein with multiple splicing RBPMS 1.59 5E−07 213577_at squalene epoxidase SQLE 1.59 8E−05 201307_at septin 11 11-Sep 1.59 9E−13 205911_at parathyroid hormone receptor 1 PTHR1 1.59 4E−05 222258_s_at SH3-domain binding protein 4 SH3BP4 1.58 2E−06 212667_at “secreted protein, acidic, cysteine-rich (osteonectin)” SPARC 1.58 1E−07 209576_at “guanine nucleotide binding protein (G protein), GNAI1 1.58 5E−07 alpha inhibiting activity polypeptide 1” 203038_at “protein tyrosine phosphatase, receptor type, K” PTPRK 1.58 3E−10 201723_s_at UDP-N-acetyl-alpha-D-galactosamine: polypeptide N- GALNT1 1.58 3E−08 acetylgalactosaminyltransferase 1 (GalNAc-T1) 220174_at hypothetical protein FLJ23420 FLJ23420 1.58 6E−05 211592_s_at NA NA 1.58 1E−05 216269_s_at “elastin (supravalvular aortic stenosis, ELN 1.58 5E−05 Williams-Beuren syndrome)” 1554195_a_at NA NA 1.58 2E−05 226408_at TEA domain family member 2 TEAD2 1.58 2E−05 227998_at S100 calcium binding protein A16 S100A16 1.57 2E−07 241355_at Hairless homolog (mouse) HR 1.57 3E−07 200755_s_at calumenin CALU 1.57 4E−07 209373_at BENE protein BENE 1.57 3E−05 218176_at “melanoma antigen family F, 1” MAGEF1 1.57 3E−10 213230_at paraneoplastic antigen HUMPPA 1.57 3E−06 225731_at KIAA1223 protein KIAA1223 1.57 2E−09 204976_s_at “Alport syndrome, mental retardation, midface hypoplasia and AMMECR1 1.56 1E−11 elliptocytosis chromosomal region, gene 1” 205635_at huntingtin-associated protein interacting protein (duo) HAPIP 1.56 1E−05 205020_s_at ADP-ribosylation factor-like 4A ARL4A 1.56 6E−05 201655_s_at Heparan sulfate proteoglycan 2 (perlecan) HSPG2 1.56 5E−06 210830_s_at paraoxonase 2 PON2 1.56 2E−06 231420_at gametogenetin GGN 1.56 8E−05 201722_s_at UDP-N-acetyl-alpha-D-galactosamine: polypeptide N- GALNT1 1.56 4E−08 acetylgalactosaminyltransferase 1 (GalNAc-T1) 235723_at basonuclin 2 BNC2 1.56 1E−06 212951_at G protein-coupled receptor 116 GPR116 1.56 2E−07 227295_at IKK interacting protein IKIP 1.55 5E−11 204518_s_at peptidylprolyl isomerase C (cyclophilin C) PPIC 1.55 7E−08 203797_at visinin-like 1 VSNL1 1.55 3E−06 206236_at G protein-coupled receptor 4 GPR4 1.55 7E−06 209087_x_at melanoma cell adhesion molecule MCAM 1.55 4E−05 219145_at latrophilin 1 LPHN1 1.55 5E−08 210300_at RAS (RAD and GEM)-like GTP-binding 1 REM1 1.55 4E−07 220027_s_at Ras interacting protein 1 RASIP1 1.55 1E−05 203060_s_at 3′-phosphoadenosine 5′-phosphosulfate synthase 2 PAPSS2 1.55 2E−05 203488_at latrophilin 1 LPHN1 1.54 2E−07 202515_at “discs, large homolog 1 (Drosophila)” DLG1 1.54 3E−08 221767_x_at High density lipoprotein binding protein (vigilin) HDLBP 1.54 8E−07 1553530_a_at “integrin, beta 1 (fibronectin receptor, beta polypeptide, ITGB1 1.54 2E−06 antigen CD29 includes MDF2, MSK12)” 222547_at mitogen-activated protein kinase kinase kinase kinase 4 MAP4K4 1.54 2E−09 217904_s_at beta-site APP-cleaving enzyme 1 BACE1 1.54 3E−05 236261_at Oxysterol binding protein-like 6 OSBPL6 1.54 1E−06 218162_at olfactomedin-like 3 OLFML3 1.53 8E−06 204992_s_at profilin 2 PFN2 1.53 2E−09 207034_s_at GLI-Kruppel family member GLI2 GLI2 1.53 4E−06 220778_x_at “sema domain, transmembrane domain (TM), and cytoplasmic domain, (semaphorin) 6B” SEMA6B 1.53 3E−06 1554127_s_at methionine sulfoxide reductase B3 MSRB3 1.53 1E−07 221529_s_at plasmalemma vesicle associated protein PLVAP 1.53 1E−06 203558_at cullin 7 CUL7 1.53 9E−05 203744_at high-mobility group box 3 HMGB3 1.53 2E−06 204517_at peptidylprolyl isomerase C (cyclophilin C) PPIC 1.53 8E−10 213249_at F-box and leucine-rich repeat protein 7 FBXL7 1.53 8E−08 222462_s_at beta-site APP-cleaving enzyme 1 BACE1 1.52 8E−11 217975_at WW domain binding protein 5 WBP5 1.52 1E−09 200770_s_at “laminin, gamma 1 (formerly LAMB2)” LAMC1 1.52 2E−05 202975_s_at Rho-related BTB domain containing 3 RHOBTB3 1.52 3E−05 219076_s_at “peroxisomal membrane protein 2, 22 kDa” PXMP2 1.52 2E−05 202828_s_at matrix metalloproteinase 14 (membrane-inserted) MMP14 1.51 2E−05 228327_x_at “Meis1, myeloid ecotropic viral integration site 1 MEIS3 1.51 7E−05 homolog 3 (mouse)” 203810_at “DnaJ (Hsp40) homolog, subfamily B, member 4” DNAJB4 1.51 8E−05 204619_s_at chondroitin sulfate proteoglycan 2 (versican) CSPG2 1.51 1E−05 227660_at anthrax toxin receptor 1 ANTXR1 1.51 3E−05 201876_at paraoxonase 2 PON2 1.51 9E−07 213959_s_at KIAA1005 protein KIAA1005 1.51 2E−05 226911_at hypothetical protein FLJ39155 FLJ39155 1.51 1E−06 207829_s_at BCL2/adenovirus E1B 19 kDa interacting protein 1 BNIP1 1.51 1E−05 223237_x_at “adaptor-related protein complex 2, alpha 1 subunit” AP2A1 1.51 3E−05 225688_s_at “pleckstrin homology-like domain, family B, member 2” PHLDB2 1.51 4E−05 204556_s_at DAZ interacting protein 1 DZIP1 1.51 3E−09 244126_at peroxisomal biogenesis factor 11 gamma PEX11G 1.50 4E−08 227569_at ligand of numb-protein X 2 LNX2 1.50 4E−09 225202_at Rho-related BTB domain containing 3 RHOBTB3 1.50 3E−05 218894_s_at mago-nashi homolog FLJ10292 1.50 1E−06 

What is claimed is:
 1. A method of treating a subject with rheumatoid arthritis, comprising the steps of: (a) obtaining a biological sample from the subject; (b) measuring in the biological sample expression of a combination of proteins comprising ICAM1 and CXCL13 in an immunoassay using monoclonal antibodies that specifically bind to the ICAM1 protein and the CXCL13 protein; (c) determining whether the expression of the proteins is elevated in the biological sample relative to a control sample; and (d) administering an effective amount of a rheumatoid arthritis (RA) therapeutic agent to treat the subject based on whether the expression of the ICAM1 protein or the CXCL13 protein is elevated, wherein the RA therapeutic agent is a tumor necrosis factor-alpha inhibitor or an anti-interleukin 6 receptor agent.
 2. The method of claim 1, wherein step (c) comprises determining that expression of the ICAM1 protein is elevated.
 3. The method of claim 2, wherein the ICAM1 protein is sICAM1.
 4. The method of claim 1, wherein the biological sample is serum.
 5. The method of claim 1, wherein the immunoassay is an ELISA.
 6. The method of claim 1, wherein step (c) comprises determining that expression of the CXCL13 protein is elevated.
 7. The method of claim 1, wherein step (c) comprises determining that expression of the ICAM1 protein is not elevated.
 8. The method of claim 1, wherein step (c) comprises determining that expression of the CXCL13 protein is not elevated.
 9. The method of claim 1, wherein step (d) comprises administering an effective amount of the tumor necrosis factor-alpha inhibitor.
 10. The method of claim 1, wherein step (d) comprises administering an effective amount of the anti-interleukin 6 receptor agent.
 11. A method of measuring protein expression, comprising the steps of: (a) obtaining a serum sample from a subject with rheumatoid arthritis; and (b) measuring in the biological sample expression of a combination of proteins comprising ICAM1 and CXCL13 in an immunoassay using monoclonal antibodies that specifically bind to the ICAM1 protein and the CXCL13 protein.
 12. The method of claim 11, wherein the immunoassay is an ELISA.
 13. The method of claim 11, wherein the ICAM1 protein is sICAM1.
 14. The method of claim 11, further comprising determining that expression of the ICAM1 protein is elevated.
 15. The method of claim 11, further comprising determining that expression of the CXCL13 protein is elevated.
 16. The method of claim 11, further comprising determining that expression of the ICAM1 protein is not elevated.
 17. The method of claim 11, further comprising determining that expression of the CXCL13 protein is not elevated. 